Switching from membrane disrupting to membrane crossing, an effective strategy in designing antibacterial polypeptide.

Drug-resistant bacterial infections have caused serious threats to human health and call for effective antibacterial agents that have low propensity to induce antimicrobial resistance. Host defense peptide-mimicking peptides are actively explored, among which poly-ß-l-lysine displays potent antibacterial activity but high cytotoxicity due to the helical structure and strong membrane disruption effect. Here, we report an effective strategy to optimize antimicrobial peptides by switching membrane disrupting to membrane penetrating and intracellular targeting by breaking the helical structure using racemic residues. Introducing ß-homo-glycine into poly-ß-lysine effectively reduces the toxicity of resulting poly-ß-peptides and affords the optimal poly-ß-peptide, ßLys50HG50, which shows potent antibacterial activity against clinically isolated methicillin-resistant Staphylococcus aureus (MRSA) and MRSA persister cells, excellent biosafety, no antimicrobial resistance, and strong therapeutic potential in both local and systemic MRSA infections. The optimal poly-ß-peptide demonstrates strong therapeutic potential and implies the success of our approach as a generalizable strategy in designing promising antibacterial polypeptides.

Biodegradable peptide polymers as alternatives to antibiotics used in aquaculture.

The pressure of antimicrobial resistance has forced many countries to reduce or even prohibit the use of antibiotics in feed. Therefore, it is an urgent need to develop alternatives to antibiotics to control infectious diseases in feed and aquaculture. To address this long-lasting challenge, we prepared peptide polymers that display potent and broad-spectrum activity against common pathogenic bacteria in aquaculture, low hemolysis and low cytotoxicity, and do not induce bacteria to develop resistance or cross-resistance to antibiotics. The optimal peptide polymer demonstrates strong in vivo therapeutic potential in an adult zebrafish infection model. Moreover, the optimal peptide polymer is biodegradable by enzymes into single amino acids and dipeptides to totally lose its antibacterial activity and, therefore, will not cause antimicrobial selective pressure. Our study suggests that peptide polymers are promising alternatives to antibiotics in aquaculture and open new avenues to address the global challenge of antimicrobial resistance.

The membrane-targeting mechanism of host defense peptides inspiring the design of polypeptide-conjugated gold nanoparticles exhibiting effective antibacterial activity against methicillin-resistant Staphylococcus aureus.

Multidrug-resistant bacterial infections are a grand challenge to global medical and health systems. Therefore, it is urgent to develop versatile antibacterial strategies that can combat bacterial resistance without displaying toxicity. Here, we synthesize antibacterial polypeptide-conjugated gold nanoparticles that exhibit potent antibacterial activities against clinically isolated multiple drug resistance Gram-positive bacteria, such as methicillin-resistant Staphylococcus aureus, and excellent in vitro and in vivo biocompatibility. The antibacterial mechanism study indicates that over-production of reactive oxygen species results in the killing of bacteria. The overall antibacterial performance of these polypeptide-conjugated gold nanoparticles and the convenient synthesis of these polypeptides via lithium hexamethyldisilazide-initiated fast ring-opening polymerization on α-amino acid N-carboxyanhydride imply the potential application of this strategy in treating bacterial infections.

Effective and biocompatible antibacterial surfaces via facile synthesis and surface modification of peptide polymers.

It is an urgent need to tackle drug-resistance microbial infections that are associated with implantable biomedical devices. Host defense peptide-mimicking polymers have been actively explored in recent years to fight against drug-resistant microbes. Our recent report on lithium hexamethyldisilazide-initiated superfast polymerization on amino acid N-carboxyanhydrides enables the quick synthesis of host defense peptide-mimicking peptide polymers. Here we reported a facile and cost-effective thermoplastic polyurethane (TPU) surface modification of peptide polymer (DLL: BLG = 90 : 10) using plasma surface activation and substitution reaction between thiol and bromide groups. The peptide polymer-modified TPU surfaces exhibited board-spectrum antibacterial property as well as effective contact-killing ability in vitro. Furthermore, the peptide polymer-modified TPU surfaces showed excellent biocompatibility, displaying no hemolysis and cytotoxicity. In vivo study using methicillin-resistant Staphylococcus aureus (MRSA) for subcutaneous implantation infectious model showed that peptide polymer-modified TPU surfaces revealed obvious suppression of infection and great histocompatibility, compared to bare TPU surfaces. We further explored the antimicrobial mechanism of the peptide polymer-modified TPU surfaces, which revealed a surface contact-killing mechanism by disrupting the bacterial membrane. These results demonstrated great potential of the peptide-modified TPU surfaces for practical application to combat bacterial infections that are associated with implantable materials and devices.

Bio-inspired poly-DL-serine materials resist the foreign-body response.

Implantation-caused foreign-body response (FBR) is a commonly encountered issue and can result in failure of implants. The high L-serine content in low immunogenic silk sericin, and the high D-serine content as a neurotransmitter together inspire us to prepare poly-DL-serine (PSer) materials in mitigating the FBR. Here we report highly water soluble, biocompatible and easily accessible PSer hydrogels that cause negligible inflammatory response after subcutaneous implantation in mice for 1 week and 2 weeks. No obvious collagen capsulation is found surrounding the PSer hydrogels after 4 weeks, 3 months and 7 months post implantation. Histological analysis on inflammatory cytokines and RNA-seq assay both indicate that PSer hydrogels show low FBR, comparable to the Mock group. The anti-FBR performance of PSer hydrogels at all time points surpass the poly(ethyleneglycol) hydrogels that is widely utilized as bio-inert materials, implying the potent and wide application of PSer materials in implantable biomaterials and biomedical devices.

Host Defense Peptide Mimicking Peptide Polymer Exerting Fast, Broad Spectrum, and Potent Activities toward Clinically Isolated Multidrug-Resistant Bacteria.

Multidrug-resistant (MDR) bacteria have emerged quickly and have caused serious nosocomial infections. It is urgent to develop novel antimicrobial agents for treating MDR bacterial infections. In this study, we isolated 45 strains of bacteria from hospital patients and found shockingly that most of these strains were MDR to antimicrobial drugs. This inspired us to explore antimicrobial peptide polymers as synthetic mimics of host defense peptides in combating drug-resistant bacteria and the formidable antimicrobial challenge. We found that peptide polymer 80:20 DM:Bu (where DM is a hydrophilic/cationic subunit and Bu is a hydrophobic subunit) displayed fast bacterial killing, broad spectrum, and potent activity against clinically isolated strains of MDR bacteria. Moreover, peptide polymer 80:20 DM:Bu displayed potent in vivo antibacterial efficacy, comparable to the performance of polymyxin B, in a Pseudomonas aeruginosa (P. aeruginosa) infected rat full-thickness wound model. The peptide polymer can be easily synthesized from ring-opening polymerization with remarkable reproducibility in structural properties and biological activities. The peptide polymer's potent and broad spectrum antimicrobial activities against MDR bacteria in vitro and in vivo, resistance to proteolysis, and high structural diversity altogether imply a great potential of peptide polymer 80:20 DM:Bu in antimicrobial applications as synthetic mimics of host defense peptides.

An alpha/beta chimeric peptide molecular brush for eradicating MRSA biofilms and persister cells to mitigate antimicrobial resistance.

Infections involving methicillin-resistant Staphylococcus aureus present great challenges, especially when biofilms and persister cells are involved. In this work, an α/ß chimeric polypeptide molecular brush (α/ß CPMB) is reported to show excellent performance in inhibiting the formation of biofilms and eradicating established biofilms. Additionally, the polymer brush efficiently killed metabolically inactive persister cells that are antibiotic-insensitive. Antimicrobial mechanism studies showed that α/ß CPMB causes membrane disturbance and a substantial increase in reactive oxygen species (ROS) levels to kill bacteria, and mesosome-like structure formation was also observed. Furthermore, the polymer brush was able to kill clinically isolated multidrug resistant Gram-positive bacteria with no risk of antimicrobial resistance. The α/ß CPMB has demonstrated great potential in addressing the great challenge of eradicating multidrug resistant Gram-positive bacterial infections.

Host defense peptide mimicking poly-ß-peptides with fast, potent and broad spectrum antibacterial activities.

Microbial infections have always been serious challenges to human health considering that antibiotics almost inevitably induce microbial resistance. Therefore, it is urgent to develop a new antibacterial agent that is active against drug-resistant bacteria and is less susceptible to microbial resistance. In this work, a series of host defense peptide (HDP) mimicking antibacterial poly-ß-peptides were synthesized, characterized and evaluated for their biological activities. The best poly-ß-peptide within this study (20 : 80 Bu : DM) displays potent and broad spectrum antibacterial activity against antibiotic-resistant super bugs and low toxicity toward mammalian cells. Moreover, these poly-ß-peptides are bactericidal and kill bacteria very fast within 5 min. An antimicrobial resistance test demonstrated that bacteria develop no resistance toward the selected poly-ß-peptides even over 1000 generations. Our studies demonstrate that random copolymers of heterochiral poly-ß-peptides, without the need for defined secondary structures, can mimic the antimicrobial HDP. These results imply the potential application of these poly-ß-peptides as new antimicrobial agents to tackle drug resistant antimicrobial infections.

[Study on antibacterial properties of titanium metallic surface due to synergistic action of micro/nano-structure and antimicrobial peptides].

Objective: To investigate the effects of micro/nano-structure and antimicrobial peptides (AMPs) on antibacterial properties for titanium (Ti) metal surface. Methods: Ti disks were treated via sandblasted large-grit acid-etched (SLA) and alkali-heat treatment (AHT) to build the micor/nano-structure, on which AMPs were spin-coated with a certain amount (10, 30, 50, 70, and 90 µg). Scanning electron microscope (SEM) and energy dispersive spectroscopy (EDS) were used to observe the surface structure and characterize the surface elements (i.e. contents of C, N, O, and Ti). Ti disks loaded with AMPs of difference amounts were co-cultured with Staphylococcus aureus ( S. aureus) for 24 hours. After that, the formation and dimension of antibacterial circle were measured. Furthermore, the Ti disks treated with different approaches (untreated, SLA treatment, SLA+THA treatment, and loaded with 90 µg AMPs) were co-cultured with S. aureus and Escherichia coli ( E.coli) for 3 hours, bacterial adhesion on the disks were evaluated by using SEM. The antibacterial performances in solution were quantitatively evaluated by immersing the Ti disks in bacterial solutions and measuring the absorbance ( A) values. Results: It was found that the nanoporous structure could be easily constructed by SLA+AHT approach. After spin-coating AMPs, the nanopores with the diameter less than 200 nm were almost covered. According to the element analysis, with the increase of AMPs, the C content gradually increased; the N content was not detected until AMPs amount reached 70 µg on the disks. The diameter of antibacterial circle clearly depended on the AMPs amount. The Ti disks loaded with 90 µg AMPs had significantly larger antibacterial circles than the other Ti disks ( P<0.05). Based on the SEM observation, the Ti disks loaded with 90 µg AMPs has the least bacterial attachment compared with the other Ti disks ( P<0.05). The A value of bacterial solution immersed with the Ti disks loaded with 90 µg AMPs was much lower than the other Ti disks ( P<0.05). Conclusion: The approach of micro/nano-structure and AMPs can improve the antibacterial properties of Ti metal surface.

Surface Modified with a Host Defense Peptide-Mimicking ß-Peptide Polymer Kills Bacteria on Contact with High Efficacy.

Methicillin-resistant Staphylococcus aureus (MRSA) has been one of the major nosocomial pathogens to cause frequent and serious infections that are associated with various biomedical surfaces. This study demonstrated that surface modified with host defense peptide-mimicking ß-peptide polymer, has surprisingly high bactericidal activities against Escherichia coli ( E. coli) and MRSA. As surface-tethered ß-peptide polymers cannot move freely to adopt the collaborative interactions with bacterial membrane and are too short to penetrate the cell envelop, we proposed a mode of action by diffusing away the cell membrane-stabilizing divalent ions, Ca2+ and Mg2+. This hypothesis was supported by our study that Ca2+ and Mg2+ supplementation in the assay medium causes up to 80% loss of bacterial killing efficacy and that the addition of divalent ion chelating ethylenediaminetetraacetic acid into the above assay medium leads to significant recovery of the bacterial killing efficacy. In addition to its potent bacterial killing efficacy, the surface-tethered ß-peptide polymer also demonstrated excellent biocompatibility by displaying no hemolysis and supporting mammalian cell adhesion and growth. In conclusion, this study demonstrated the potential of ß-peptide polymer-modified surface in addressing nosocomial infections that are associated with various surfaces in biomedical applications.