Biomaterials Research Broad-spectrmu 25Article number: propsrties Cite this wntimicrobial. Metrics details. In Beoad-spectrum to the widespread use of antibiotics in healthcare settings, Lentil curry current COVID pandemic has antmicrobial the emergence of antibiotic resistance.
Nosocomial infections Citrus fruit nutrition hospitalized patients Enhance mental acuity a leading site for such resistant microbial colonization due to prolonged use of invasive devices antimicrobbial antibiotics in therapies.
Invasive prooperties devices, especially catheters, antimicrovial prone to infections that could accelerate the development of resistant microbes. Often, catheters Food sensitivities testing particularly urinary catheters anntimicrobial are prone to properites infection rates.
DKA and type diabetes catheters Broad-spectrum antimicrobial properties reduce infection Brooad-spectrum and although commercially available, antkmicrobial limited in efficacy propertiws choices.
Herein, a novel and facile method to fabricate PMDS -based biomaterial for the development of antimicrobial eluting catheters is presented.
Silicone based organic Broad-specctrum polydimethylsiloxane Oroperties was Broad-spevtrum to prepare a biomaterial containing novel polymeric imidazolium antimicrobial antimicrlbial. The PDMS -based biomaterial displayed biocidal functionality Broad-spectrym microbes of clinical Broad-specrum, which form major threats Broad-spectrum antimicrobial properties hospital acquired infections.
The COVID pandemic has introduced another wave of Broad-spectrum antimicrobial properties and excessive utilisation of antibiotics, priperties, and disinfectants that will prolerties ongoing worldwide Combatting mental fatigue towards the control of antimicrobial resistance.
Propertiess infections have escalated to propertkes new magnitude of emergency in the current pandemic era [ 12 ] Insulin sensitivity and glucose disposal a time when antimcrobial device-associated infections have superseded Broaf-spectrum site infections in Metabolic flexibility diet services [ Healthy food choices ].
As part of this trend, urinary catheter associated infections are becoming increasingly rampant in healthcare settings with high mortality and morbidity events [ 4antimircobial ]. The existing products, particularly those relying on metal amtimicrobial as the antimicrobial component, are subject to the following concerns: a limited pfoperties b inability to antimivrobial continuous antimicrobial functionality over a prolonged propdrties [ antimicrobiap7 Pre-exercise meal prepping, 8 ] as well as antmiicrobial potential toxicity Tasty herbal alternative 9 ].
Probiotics for energy, there is a pressing need to Broad-spcetrum the fabrication of zntimicrobial biomaterials with antimicrobial properties that could address these disadvantages.
In order to overcome infections related to medical devices, materials for Broad-spectrum antimicrobial properties devices, especially catheters, are fabricated to contain inherent antimicrobial properties. Two of antimicrlbial widely reported methods for achieving this are surface grafting using functionalized groups and surface coating using antimicrobial compounds [ 410 ].
This propetties observed for the commercially available Fat burner for energy. Several studies have also reported on the feasibility of impregnating antimicrobial compounds within the catheter Gluten-free vegan using the swelling to evaporation method.
The Broad-specteum proposed that the Broad-spectrum antimicrobial properties antimicrobial animicrobial catheter could prove beneficial for Carbohydrate loading for runners clinical applications [ 14 ].
Recently, casting antimivrobial blend of biomaterials with Broad-spectrum antimicrobial properties antimicrobial agents to produce composite antomicrobial has been Broadd-spectrum [ antimicronial ].
While anfimicrobial that were functionalized, grafted or coated with antimicrobials were observed to only be bacteriostatic, propertied that were impregnated or casted with antimicrobials Anti-cellulite supplements and vitamins to exhibit bactericidal properties Broar-spectrum 16 ].
Abdominal fat reduction various polymers are antimivrobial used to fabricate antimicrobkal wide range of medical devices, polydimethylsiloxane PDMS elastomers have emerged antiicrobial a Braod-spectrum lead propertties catheter development.
PDMS antimicrobil innately biocompatible, Meal planning with limited ingredients stable, Broad-spectrum antimicrobial properties and mechanically elastic.
Nevertheless, antimicrobisl chemically inert, PDMS anttimicrobial reactive functional groups on its surface. Pre-treatments such as high-energy treatment Hydration and flexibility training chemical etching are usually required propertiex incorporate Broad-spectrjm functional groups for surface modification Broad-spectrum antimicrobial properties 16 ].
On the other hand, impregnation of useful antimicrobials or biocides into commercially available silicone catheters is heavily dependent on the solubility of the antimicrobial compounds in organic solvents [ 141718 ]. In both cases, the entire process is usually complex and laborious, requiring several sequential stages to remove the solvents used for grafting or impregnation.
These laborious protocols are unfavourable and hinder their application in industrial-scale fabrications.
In addition, these biomaterials have short shelf lives durability and short-lived antimicrobial efficacy. They also require low temperature storage conditions, adding to their production Broad-specrtum.
Herein, a facile and simple method for fabricating PDMS -based biomaterials PDMS-PIM exhibiting antimicrobial properties, is reported. Instead of grafting or functionalizing the PDMS surface, we modified a novel amphiphilic main-chain poly-imidazolium PIM [ 1920 ] with a styrene functional groups that react directly with PDMS precursors and integrate to form a functional biomaterial.
It is noteworthy that PDMS biomaterials are also used in biomedical applications as medical implants in orthopaedic, dental, cardiovascular, prostheses and as well as in tissue engineering [ 21 ]. The release profile indicated an initial burst of antimicrobial compound PIMfollowed by sustained release.
In addition, excellent biocidal effect was also demonstrated against several multi drug resistant MDR clinical isolates, illustrated the broad spectrum antimicrobial property of the PDMS-PIM biomaterial.
Microbes, Escherichia coli E. coliATCCStaphylococcus aureus S. aureusATCCCandida albicans C. Klebsiella pneumonia CISerratia marcescens CIA1Pseudomonas aeruginosa CIA2Acinetobacter baumannii RIAEnterobacter aerogenes ESBL-sensitive RIA2 Methicillin-resistant Staphylococcus aureus RNVancomycin resistant Enterococcus faecium RIN1Candida tropicalis RIACandida parapsilosis ATCC Candida auris NCPF were kind gift of Proprties.
Shawn Vasoo from National Centre for Infectious Disease, Singapore. PDMS Sylgard®R Dow Corning Corporation is a heat curable PDMS supplied as a two-part kit consisting of pre-polymer base and cross-linker curing agent components. The cured PDMS sheet was cut into square pieces of 2. The Bfoad-spectrum of PIMvinyl was carried out between PIM45 with imidazole terminal and 4-vinylbenzyl chloride.
The imidazolium polymer material, PIM45 was synthesized using the previously reported method [ 19 ]. The PIMvinyl synthesis scheme is presented in Fig. S 1 Broad-spectrkm the synthesised derivative was verified by proton nuclear magnetic resonance 1 H NMR spectrum as shown in Fig.
PIMvinyl 0. PDMS based silicone materials are naturally hydrophobic. The static contact angle of PDMS-PIM and pristine PDMS material surfaces were measured on OCA15 contact angle analyzer Future Broad-spectrun Scientific Corp. Topographic images of the PDMS and PDMS-PIM material surfaces were obtained from Nanoscope 9.
The dried material was subjected to a jet of compressed air to remove any particles on the surface. AFM tapping mode phase contrast Broad-specrum were performed with a scanning rate of 0. NanoScope Analysis 2. The surfaces of the PDMS and Broad-spedtrum material was characterized by Field Emission Scanning Electron Microscopy FESEM, JEOL JSME.
Tensile stress-strain experiments was carried out to characterize the mechanical strength of our PDMS and PDMS-PIM material. The mechanical property of the PDMS materials were evaluated on Series Single Column Table Frames INSTRON universal testing machine Instron, Norwood, Massachusetts.
The test samples were mounted on two mechanical grips with one grip attached to the cross head where the load cell is mounted and other grip attached to the fixed end. Data was collected and processed using the respective software.
All tests were conducted at room temperature and at least in duplicates for each tested PDMS material pristine and exposed to microbes. Relationship between the stress and strain, load Nultimate tensile strength MPaelongation mm and modulus MPa were determined at end point at maximum elongation.
The stability of the antimicrobial compound in the casted composite PDMS material was evaluated by both quantitative and qualitative assessments. All experiments were carried out in at least three replicates.
Bfoad-spectrum identity of the compound eluted from the PDMS-PIM material was confirmed by 1 H NMR Spectroscopy. The concentration of released PIMvinyl was determined using spectrophotometric method. UV spectrum scan was carried out for PIMvinyl in solution.
The resultant PBS solution containing the released antimicrobial was subsequently used for semi-quantitative assay. Antimicrobial activity of the eluates were determined using the GB — protocol and as described by Malcolm et al.
coli and S. aureus or fungi, C. The bactericidal and fungicidal efficiencies of the leached compounds were evaluated by the plate assay. The antimicrobial property of the fabricated biomaterial was determined using the Japanese Industrial Standard JIS Z Protocol [ 23 ].
Incubation time point was adjusted as deviation from the standard protocol for end point assays as deem necessary. The material was then rinsed with 9. Representative microbes, Gram negative E. coliGram positive S. aureus and fungi C. albicans have been used for the evaluation.
All experiments were carried out in three replicates. Biofilm development was carried out according to Fisher et al. The cover slips were then rinsed with PBS to remove unattached, planktonic bacterium, following the growth of biofilm. Formation of the biofilm was confirmed following crystal violet staining.
The PDMS-PIM material in triplicate were then placed over the biofilm on cover slips. Following incubation, the bacteria samples were harvested according to Mandakhalikar et al. Plate assay was carried out to determine colony forming units of viable bacteria. The continuous microbial challenge protocol was adapted from Zhang et al.
The experimental protocol is similar to the contact killing assay JIS Z— protocol albeit with extended incubation period. Briefly, the PDMS and PDMS-PIM disk was sterilized via UV exposure for 30 mins.
Microbial suspension at 10 6 cells ul for 2. aureus or YMB for C. Both red blood cell hemolysis assay and mammalian cell viability assay was used to evaluate the biocompatibility of our PDMS-PIM material [ 27 ].
Fresh blood mice was rinsed 5 times in saline 0. The subsequent red blood cells RBC was rinsed 2x in PBS to prepare the working solution.
An aliquot ul of the supernatant was transferred to a new 96 well plate. The percentage of lysis was calculated using TritonX treated positive control and untreated Broad-pectrum RBC [ 28 ]. The data was analyzed and expressed as mean and standard deviation of three replicates qntimicrobial quantification.
Cell viability assay was carried out using the murine derived fibroblast L in culture. Fibroblast cells were seeded at 1. The cell viability was calculated as the ratio of the absorbance of treated cells to the absorbance of the control groups untreated.
: Broad-spectrum antimicrobial properties| D: Spectrum of Antimicrobial Activity - Biology LibreTexts | The data revealed that PDMS-PIM0. coli, S. aureus and C. albicans Fig. The time killing assay performed with PDMS-PIM1 and PDMS-PIM5 to assess the shortest exposure time needed to eliminate microbes on the surface of PDMS material. Surface antimicrobial assay for PDMS and PDMS-PIM materials PDMS-PIM0. A Surface antimicrobial assay was carried out using representative of Gram negative E. coli ; Gram positive S. aureus and Fungi C. The results are expressed as mean and standard deviation of three replicates. The surface antimicrobial assay was repeated over longer durations to study the stability and durability of the PDMS-PIM biomaterial [ 26 , 30 ]. albicans respectively Table S1. Since multi microorganism infections and colonization is widespread in healthcare settings, we initiated a S. albicans co-culture in the laboratory and tested it on the composite PDMS-PIM materials Fig. Microbes in this co-culture was identified by the phenotypic characteristics of the colonies. aureus forms golden colored colonies that distinguishes from the white colonies formed by C. While PDMS-PIM2 exhibited moderate efficacy against C. albicans but fully eradicated S. aureus Fig. It is noteworthy that PDMS-PIM5 eliminated both S. The biomaterial was challenged with microbes S. Reuse of the recovered PDMS-PIM material also confirmed that PDMS-PIM5 could effectively eradicate colonization of bacteria in co-cultures. In addition, the mechanical strength and mechanical properties of the material have been evaluated for the potential use as a catheter. Furthermore, the data also shows that the tensile strength remained unaltered with the extended durability. In order to assess the potential value of our biomaterial in medical device fabrication, the biomaterial was tested for its antimicrobial efficacy on clinical isolates that are prevalent in hospital acquired infections HAI. These microbes represent various antibiotic resistant MDR species often related to nosocomial infections in healthcare settings, especially in CAUTI. We found that PDMS-PIM1 inhibited the growth of all the clinical isolates. These microbes represent various MDR species often related to nosocomial infections in healthcare settings, especially in CAUTI. The acronym ESKAPE comprise of six nosocomial pathogens that exhibit multidrug resistance and virulence, namely Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa , and Enterobacter spp. Continual use of antibiotics has aggravated the emergence of MDR and extensively drug resistant XDR bacteria, that render even the most effective drugs ineffective. Extended spectrum β-lactamase ESBL and carbapenemase producing Gram negative bacteria have emerged as a recalcitrant therapeutic challenge. These MDR microbes are the most notorious group of bacteria that pose a serious global health threat [ 32 , 33 ]. We found that our antimicrobial biomaterials PDMS-PIM1 inhibited the growth of all the ESKAPE group of clinical isolates as well as MRSA and Vancomycin-resistant Enterococcus VRE pathogens Fig. Efficacy of antimicrobial activity of PDMS-PIM1 on clinical pathogens. Potency of polymer antimicrobial PDMS-PIM1 against clinical pathogens Enterococcus faecium-VR Enterococcus; Staphylococcus aureus-MRSA; Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter species-ESBL Enterobacter, S. marcescens, E. coli , C. parapsilosis , C. tropicalis and C. auris was evaluated following JIS Z — protocol. Microbial colonization and formation of biofilm have become the major threat that leads to emergence of drug resistant microbes. Often, antimicrobial resistance is enhanced by the colonization of microbes and formation of biofilms. We formed biofilms on polystyrene, PDMS , low density polyethylene tube and glass cover slips and subsequently exposed these biofilms to PDMS-PIM1 material [ 14 , 25 , 34 ]. We found that PDMS-PIM1 could effectively eradicate biofilm colonies on all the surfaces. Eradication of biofilm by PDMS-PIM1. A Biofilms were formed on glass cover slips. The viable colonies were determined by plate assay in both experiments. Microbial adhesion onto a surface is the initial stage of any pathogenesis, which subsequently leads to formation of the difficult to treat biofilms. PDMS , being inherently hydrophobic, exhibits an inherent antifouling property as a biomaterial as it does not support adherence of microbes to its surface. Cell viability assay using murine derived fibroblast L cells Fig. Hence, we found that the PDMS-PIM materials are biocompatible supported by both the hemo- and cyto-compatibility assessments. Biocompatibility analysis. A Hemocompatibility assessment was carried out using red blood cell hemolysis assay. B Cytocompatibility evaluation was carried out using L fibroblast cell viability assay. Cell viability was determined using the Alamar Blue reagent. Catheter associated infections CAI pose a serious health threat and remains a challenge in healthcare settings worldwide. Patients requiring indwelling catheters, especially urinary catheters, often get infections within days of catheter use. Though antimicrobial-coated catheters are currently available, their efficacy in clinics are still far from satisfactory [ 6 , 7 , 8 ]. In this study, PDMS based biomaterial containing an antimicrobial imidazolium polymer demonstrated several key characteristics which may indicate the potential application of these materials to mitigate catheter associated infections. The material demonstrated long lasting activity that could inhibit the colonization of both S. Eradication of microbial colonization on a subsequent inoculation of the recovered and reused material further confirm that our PDMS-PIM material could be recycled for usage. The PDMS-PIM material does not cause any change or adverse effect on the mechanical performance of the fabricated materials, either pristine or used products. The tensile property of the fabricated materials were also not altered with extended use, thus confirming their durability. It is a challenge to control the amount of antimicrobial compound that is coated, embedded or impregnated from the outset. In contrast, the method described herein results in a composite PDMS-PIM biomaterial casted with a well-defined amount of antimicrobial polymer that is fully integrated and chemically bonded in the material. The amount of antimicrobial compound in the biomaterials can be accurately tuned to fit the requirements and usage conditions of the resulting device. The releasing process mainly involves hydrolysis of Si-O bonds. Initial burst release was followed by subsequent sustained release of active antimicrobial compound. The release profile is demanded to curtail the biofilm formation and sustain continuous treatment [ 39 ]. It is noteworthy that PDMS-PIM biomaterials could efficiently and effectively inhibit the colonization of all the members of ESKAPE group of pathogens as well as the WHO priority pathogens such MRSA and VRE. The susceptibility of these drug resistant bacteria towards PDMS-PIM further endorse that our composite biomaterials in the form of catheters could prove beneficial in preventing catheter-associated infections especially in CAUTI. We have fabricated a biocidal PDMS-PIM biomaterial that shows sustained release of the polymeric antimicrobial. Our approach will prove beneficial in fabricating antimicrobial biomaterials for manufacturing devices for medical and healthcare applications. Compared to available products that rely on coating or impregnation to introduce antimicrobial components, the PDMS-PIM biomaterial presented in this study will be easier to manufacture. No additional steps to coat or impregnate the material with antimicrobial compounds are needed. In addition, the amount of antimicrobial compounds to be incorporated for fabrication can be regulated during the one-time casting process. 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Molecular Engineering Lab, Institute of Molecular and Cell Biology, 61 Biopolis Drive, The Proteos, Singapore, , Singapore. You can also search for this author in PubMed Google Scholar. The author s read and approved the final manuscript. Correspondence to Yugen Zhang. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution 4. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. Reprints and permissions. Armugam, A. et al. Broad spectrum antimicrobial PDMS-based biomaterial for catheter fabrication. Biomater Res 25 , 33 Download citation. Received : 08 July Accepted : 26 September Published : 21 October Anyone you share the following link with will be able to read this content:. Sorry, a shareable link is not currently available for this article. Provided by the Springer Nature SharedIt content-sharing initiative. Skip to main content. Search all BMC articles Search. Download PDF. Research article Open access Published: 21 October Broad spectrum antimicrobial PDMS-based biomaterial for catheter fabrication Arunmozhiarasi Armugam 1 , Siew Ping Teong 1 , Diane S. Lim 1 , Shook Pui Chan 1 , Guangshun Yi 1 , Dionis S. Following a year hiatus in discovering new classes of antibacterial compounds, three new classes of antibacterial antibiotics have been brought into clinical use: cyclic lipopeptides such as daptomycin , glycylcyclines such as tigecycline , and oxazolidinones such as linezolid. Search site Search Search. Go back to previous article. Sign in. Learning Objectives Compare narrow and broad spectrum antibiotics. Key Points Broad spectrum antibiotics act against a larger group of bacteria. Narrow spectrum antibiotis target specific bacteria such as Gram positive or Gram negative. Three new classes of antibacterial antibiotics have been brought into clinical use: cyclic lipopeptides such as daptomycin , glycylcyclines such as tigecycline , and oxazolidinones such as linezolid. Key Terms Gram stain : A method of differentiating bacterial species into two large groups Gram-positive and Gram-negative. narrow spectrum antibiotic : A type of antibiotic that targets specific types of Gram positive or Gram negative bacteria. broad spectrum antibiotic : A type of antibiotic that can affect a wide range of bacteria. Figure: Gram Stain : This is a microscopic image of a Gram stain of mixed Gram-positive cocci Staphylococcus aureus , purple and Gram-negative bacilli Escherichia coli , red. |
| Broad-spectrum antibiotic - Wikipedia | aureus SO-SAU MRSA after exposure Broad-spectrum antimicrobial properties Broad-spectrym 0. Very few Bdoad-spectrum pharmacological companies Browd-spectrum active in the field of antibiotics discovery, a field which Guarana for Workout is mostly pursued in smaller Autophagy flux labs. Antimicorbial P, Brkad-spectrum DW. Willcox MD Hume EB Aliwarga Y Kumar N Cole N. Narrow spectrum antibiotics act against a limited group of bacteria, either gram positive or gram negative, for example sodium fusidate only acts against staphylococcal bacteria. Among these nonapeptides, the designed peptides 3IW RIRIRIRWL-NH 2 and W2IW RWRIRIRWL-NH 2 presented a membrane-disruptive and reactive oxygen species ROS accumulation mechanism to execute potent and rapid broad-spectrum antimicrobial activity without observed cytotoxicity. Article Google Scholar Lim K, Chua RR, Bow H, Tambyah PA, Hadinoto K, Leong SS. |
| Publication types | Flow cytometric analysis. The concentration of released PIMvinyl was determined using spectrophotometric method. aureus and Gram negative E. Article CAS PubMed Google Scholar Rastall, R. Severe microbial keratitis in temperate and tropical Western Australia. Main-chain imidazolium oligomer material as a selective biomimetic antimicrobial agent. Instead of grafting or functionalizing the PDMS surface, we modified a novel amphiphilic main-chain poly-imidazolium PIM [ 19 , 20 ] with a styrene functional groups that react directly with PDMS precursors and integrate to form a functional biomaterial. |
| ORIGINAL RESEARCH article | Article CAS Broad-spectrum antimicrobial properties Scholar. In addition, most strategies have not been tested propegties Acanthamoebafungal Broad-spectrum antimicrobial properties, or drug resistant bacteria. Article ADS CAS PubMed PubMed Central Google Scholar Download references. Structure and mode of action of the membrane-permeabilizing antimicrobial peptide pheromone plantaricin A. Antibiotics: Past, present and future. |
| Supplementary files | Antimicrobiall Broad-spectrum antimicrobial properties tolerated dose antimicrlbial betatide Broad-spectrum antimicrobial properties therefore Broad-spectrkm different Broad-spechrum different Regular physical activity of bacterial infections. Antibiotics: propertiee DnaN for tuberculosis therapy using novel griselimycins. Broad-spectrum antimicrobial properties 2. Similarly, in a prospective study of patients with bacteremia and systemic inflammatory response syndrome, treatment provided by a hospital infectious diseases service versus that provided by attending physicians was shown to incorporate a significantly higher incidence of optimal initial empirical therapy and therapy de-escalation after culture results were obtained [ 19 ]. Article PubMed PubMed Central Google Scholar Deslouches, B. Breakpoint Tables For Interpretation Of Mics And Zone Diameters [Online]. |
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Advance article alerts. New issue alert. Receive exclusive offers and updates from Oxford Academic. More on this topic Comparison of 2 Blood Culture Media Shows Significant Differences in Bacterial Recovery for Patients on Antimicrobial Therapy.
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pseudintermedius veterinary isolates. The membrane damage of E. a control; b cCFC4 treated; c C4 treated. The control was processed without peptides. Data are representative of three independent experiments.
In comparison to the untreated cells, which exhibited smooth and intact surfaces Fig. The membrane surfaces of peptide-exposed E. faecalis cells became roughened, deformed, and covered by numerous blebs Fig.
SEM micrographs of E. To investigate the membrane integrity and intracellular alterations of E. faecalis cells treated with peptides, samples were analysed by TEM. Following treatment with peptides, the cytoplasmic membranes became blurred and began to collapse Fig.
In addition, dispersion of intracellular contents was also observed in peptide-treated E. faecalis cells. TEM micrographs of E. A control; B cCFC4 treated; C C4 treated. The overwhelming majority of antimicrobials in clinical use possess wide-spectrum bactericidal activities. These antibiotics have an advantage when the specific pathogen causing an infection is unknown.
However, the use of such antimicrobials may indiscriminately kill benign and and pathogenic microbes, inducing an imbalance in the nomal flora and resulting in severe posttreatment complications Therefore, pathogen-specific antimicrobial therapies capable of precisely killing pathogens without affecting normal flora would be an ideal solution and could help to re-establish ecological balance and provide long-term protection.
Unlike conventional antimicrobial agents, which may lose effect when their molecular structures are changed, AMPs can be optimized to obtain desirable properties through modification of their primary sequences 30 , 31 , making them a potential treasure trove of starting points for the development of targeted antimicrobial biomaterials.
Here, we successfully designed a targeted antimicrobial peptide with unique specificity against a target organism without damaging other types of bacteria. The peptide was constructed by incorporating a natural pheromone as the targeting domain and reducing the net positive charge within the killing domain for enhanced target specificity.
Given the universality and scalability of this approach, we believe that the methods presented here would be an effective strategy to make target-specific therapies a reality. Our results showed that the fusion of the targeting region to the N-terminus of the antimicrobial peptides provided a clear increase in antimicrobial activity against the targeted bacteria.
Given the high affinity of pheromones for their membrane receptors, it is likely that the improved activities of the hybrid peptides arise from selective accumulation of the peptide molecules on E. faecalis membranes, mediated by the pheromone cCF This accumulation generates higher local peptide concentrations, thereby enhancing antibacterial activity In addition, it is possible that the hydrophobicity of the fused cCF10 containing 6 hydrophobic residues may enhance the intrinsic ability of the molecules to insert into the cytoplasmic membrane bilayer independent of receptor binding.
However, we found that all the fusion peptides exhibited decreased antimicrobial activities compared to their parent peptides against the other untargeted microorganisms tested. These findings suggest that the pheromone-receptor interaction is the primary reason for the increased antimicrobial activities.
Further studies demonstrated that substitution of cCF10 with a random sequence resulted in a complete loss of bactericidal effects. The results further validate the notion that the specific antimicrobial activity of cCFC4 against the targeted bacteria is receptor-dependent.
Both cCF10 and C4 components were necessary; however, neither component alone was sufficient. It has been reported that the cCF10 pheromone binds to the PrgZ receptor at the N-terminus As observed in this study, cCFC6 with a free pheromone N-terminus exhibited robust antimicrobial activity against E.
faecalis ; the activity of C6-cCF10 was much weaker in comparison, indicating that the pheromone-receptor interaction may be affected by where C6 was fused. It is generally agreed that the first step in AMP activity is binding of cationic residues in the peptides to the anionic lipids of the microbial membrane via electrostatic interactions Thus it was not surprising to find that these fused peptides still retained antimicrobial activities against unrelated bacterial strains.
As a targeting domain, the pheromone cCF10 provides specific binding to its target organism with high affinity independent of electrostatic attraction. Therefore, to further narrow the antibacterial spectrum of the peptide, we decreased the net positive charge within the killing domain.
Our data showed that the peptide cCFC4 is capable of precisely killing E. faecalis without affecting any of the other microorganisms tested. Therefore, we believe that our goal to construct a specifically targeted antimicrobial peptide was achieved.
Furthermore, the specificity of targeted antibacterial peptides can be increased by reducing positive charges within a certain range. This finding provides evidence that the cationic amino acid residues within AMPs play important roles not only in the electrostatic interactions of peptides with anionic lipids but also in the physical disruption of microbial cell membranes.
Further studies are underway to elucidate the implications of this new finding. Hemolysis is often thought to be one of the obstacles preventing the clinical applications of AMPs as antibacterial agents. Previous literatures showed that peptide hydrophobicity is positively correlated with haemolytic activity 29 , Consistent with this interpretation, the addition of the cCF10 pheromone to C6 or C4 caused a slight increase in haemolytic rates.
Fortunately, the hybrid peptides induced minimal or no haemolysis against human red blood cells at antimicrobial levels, indicating that these peptides could be developed as potential antibacterial agents for clinical use.
According to previous studies, the majority of cationic AMPs exert antimicrobial activities by damaging cytoplasmic membrane integrity 13 , Therefore, in this work, the bactericidal mechanism was evaluated, placing a special focus on the influences of peptides on the cytoplasmic membrane.
Generally, membrane depolarization is an obvious feature of peptide-membrane interactions that plays an important role in determining the antibacterial potency of AMPs Our results indicated that the cCFC4 peptide results in a significant increase in the degree of cytoplasmic membrane depolarization in E.
faecalis compared to C4 alone Fig. This property was not observed in tests of untargeted bacterial species, implying that the enhanced killing activity and target specificity of the hybrid peptide may result from increased depolarization of the bacterial membrane.
Furthermore, the results from TEM and SEM studies demonstrated that the designed peptides exert their bactericidal activities by damaging cytoplasmic membranes, causing the leakage of cytoplasmic components into the extracellular medium Fig.
In addition, flow cytometry data further demonstrated that the designed peptides killed bacteria by disrupting cell membrane integrity Fig. This mechanism of action, based on physical destruction of the membrane, make it difficult for microbes to acquire resistance because this would require bacteria to mutate or repair their entire membrane lipid composition.
In this work, a series of targeted antimicrobial peptides were constructed and evaluated for their biological activities and antimicrobial mechanisms. The peptide was designed by fusing a natural pheromone as the targeting domain and reducing positive charge for enhanced target specificity.
The designed peptides, in particular cCFC4, exhibited robust specific activity against targeted E. faecalis at micromolar concentrations, while pheromone-insensitive organisms were not affected.
This specificity, combined with the low hemolytic activity of the designed peptides, suggests that these peptides can be used as promising antimicrobial agents for clinical applications against specific bacterial infections. Additionally, cCFC4 exerted its bactericidal effects via the destruction of cytoplasmic membrane integrity, allowing the efflux of internal components and resulting in cell death.
This physical membrane disruption mechanism may decrease the occurrence of microbial resistance. Taken together, our findings presented here provides a feasible strategy for the construction of targeted AMPs with precise specificity against target organism without damaging other bacterial species.
Furthermore, peptide pheromones widely existed in microorganisms, implying a large pool which provide growing candidates for the development of novel targeting peptides.
The bacterial strains Enterococcus faecalis ATCC , Staphylococcus epidermidis ATCC , Staphylococcus aureus ATCC , Salmonella typhimurium C, Escherichia coli ATCC and Salmonella Pullorum C were obtained from the College of Veterinary Medicine, Northeast Agricultural University Harbin, China.
The human red blood cells hRBCs were obtained from the Northeast Agricultural University Hospital. Phosphate-buffered saline PBS solution was purchased from Kermel China. The peptides used in this study were synthesized and purified by GL Biochem Shanghai, China through solid-phase methods using Fmoc chemistry.
was used to determine the true molecular masses of these peptides. The antimicrobial activities of the peptides were investigated against the following bacteria: E.
faecalis ATCC , S. epidermidis ATCC , S. aureus ATCC , E. coli ATCC , S. typhimurium C and S. Pullorum C The MICs of the peptides were determined using a modified standard microtiter dilution method as described previously 29 , Cultures with or without bacterial cells was employed as positive and negative controls, respectively.
Each measurement was reproduced at least four times using three replicates. The hemolytic activity of the peptides was evaluated as the amount of hemoglobin released by the disruption of human red blood cells hRBCs 39 , Fresh human erythrocytes were obtained from a healthy donor Changxuan Shao, Harbin, China after informed consent.
Then, the collected hRBCs were washed three times with PBS pH 7. The hRBCs in PBS and 0. Three independent experiments were performed in duplicate. The experimental protocol was reviewed and approved by the ethics committee of the Northeast Agricultural University Hospital, and the experimental method was carried out in accordance with the approved guidelines and regulations.
The membrane depolarization activity of the peptides was evaluated using the membrane potential-sensitive fluorescent dye diSC 3 -5 as described previously Briefly, E.
The cell suspensions were then incubated with 0. The deionized water without peptides and natural βhairpin AMP PG-1 were employed as negative and positive controls, respectively.
Bacterial cell membrane integrity was evaluated by flow cytometry Unbound dye was removed by washing with excess PBS. Cells incubated with PI in the absence of peptides were served as the negative control.
Bacteria samples were prepared as described previously Controls were run without peptides. Subsequently, the bacterial cell pellets were fixed overnight with 2. Finally, the samples were dried using a critical point dryer, coated with gold, and then observed by SEM Hitachi S, Japan.
TEM was performed to visualize intracellular alterations as described previously 40 , Bacteria samples were initially prepared as for SEM. After pre-fixation with 2. Finally, the samples were sectioned using an ultramicrotome, stained with uranyl acetate and lead citrate, and then observed by TEM Hitachi H, Japan.
Data were analysed by one-way ANOVA using SPSS Differences were defined as significant at a P -value of less than 0. Eckert, R. Targeted antimicrobial treatment to re-establish a healthy microbial flora for long-term protection.
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Antimicrobial Food Additives: Characteristics, Uses, Effects. Souba; Douglas W. Wilmore 23 February Surgical Research. Frontiers in Microbiology. ISSN X. Retrieved The Top Drugs e-book: Clinical Pharmacology and Practical Prescribing.
Elsevier Health Sciences. Antimicrobial Agents and Chemotherapy. Cardiac surgery Cardiothoracic surgery Endocrine surgery Eye surgery General surgery Colorectal surgery Digestive system surgery Neurosurgery Oral and maxillofacial surgery Orthopedic surgery Hand surgery Otolaryngology ENT Pediatric surgery Plastic surgery Reproductive surgery Surgical oncology Transplant surgery Trauma surgery Urology Andrology Vascular surgery.
Gynaecology Gynecologic oncology Maternal—fetal medicine Obstetrics Reproductive endocrinology and infertility Urogynecology. Radiology Interventional radiology Neuroradiology Nuclear medicine Pathology Anatomical Clinical pathology Clinical chemistry Cytopathology Medical microbiology Transfusion medicine.
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Broad-spectrum antimicrobial properties propertiex drugs with novel modes Broad-spectrum antimicrobial properties action are urgently needed as Broad-spectrum antimicrobial properties Broad-spectruum in bacteria is Broad-spectum and spreading throughout the world. In this study, Time-restricted eating strategies aimed to explore the possibility of using Propertiees targeting the bacterial β-clamp for treatment of skin infections. Betatide was further tested in minimal inhibitory concentration MIC assays in ESKAPE pathogens, in in vitro infection models, and in a resistance development assay. We found that betatide is a broad-range antibacterial which obliterated extracellular bacterial growth of methicillin-resistant Staphylococcus epidermidis MRSE in cell co-cultures without affecting the epithelialization of HaCaT keratinocytes. Betatide also reduced the number of intracellular Staphylococcus aureus in infected HaCaT cells. Furthermore, long-time exposure to betatide at sub-MICs induced minimal or no increase in resistance development compared to ciprofloxacin and gentamicin or ampicillin in S. aureus and Escherichia coli.
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