Immunomodulatory Properties of Host Defence Peptides in Skin Wound Healing.

Cutaneous wound healing is a vital biological process that aids skin regeneration upon injury. Wound healing failure results from persistent inflammatory conditions observed in diabetes, or autoimmune diseases like psoriasis. Chronic wounds are incurable due to factors like poor oxygenation, aberrant function of peripheral sensory nervature, inadequate nutrients and blood tissue supply. The most significant hallmark of chronic wounds is heavily aberrant immune skin function. The immune response in humans relies on a large network of signalling molecules and their interactions. Research studies have reported on the dual role of host defence peptides (HDPs), which are also often called antimicrobial peptides (AMPs). Their duality reflects their potential for acting as antibacterial peptides, and as immunodulators that assist in modulating several biological signalling pathways related to processes such as wound healing, autoimmune disease, and others. HDPs may differentially control gene regulation and alter the behaviour of epithelial and immune cells, resulting in modulation of immune responses. In this review, we shed light on the understanding and most recent advances related to molecular mechanisms and immune modulatory features of host defence peptides in human skin wound healing. Understanding their functional role in skin immunity may further inspire topical treatments for chronic wounds.

Effects of mixed antimicrobial peptide on the growth performance, antioxidant and immune responses and disease resistance of Pengze crucian carp (Carassius auratus var. Pengze).

Antimicrobial peptides have broad-spectrum antibacterial properties and low drug resistance, and they demonstrate great potential as antibiotic substitutes. In this study, five dietary mixed antimicrobial peptide supplement groups were set and fed to Pengze crucian carp for 10 weeks. The 6 groups were G0 (control group) and 5 additional groups: G1 (100 mg/kg), G2 (200 mg/kg), G3 (400 mg/kg), G4 (800 mg/kg) and G5 (1600 mg/kg). The results showed that the final body weight (FBW), weight gain rate (WGR) and specific growth rate (SGR) of fish in G1 and G2 were higher than those of fish in the control group, and G1 was significantly higher than G0 (P < 0.05). In addition, the FBW, WGR, and SGR of the G3 group were significantly lower than those of the G0 group. The chymotrypsin, lipase and amylase activities of G1 and G2 were significantly upregulated compared with G0 and reached peak values in G1. The activity of T-AOC and SOD in the addition group was higher (except G2 and G4) than that in the control groups, and significantly increased in G3 compared to the control group. The activity of MDA in the addition group was lower than that in the control group (p > 0.05). The expression levels of TLR-4, MYD88 and TNF-α in the three organs of the addition group were higher than those in G0 and reached the peak value in G3 (p < 0.05). Furthermore, the expression levels of TLR-4, MYD88 and TNF-α in the three organs of G3 were significantly lower than those in G0 and lower than those in the other supplemented groups. The expression levels of IL-10 and IL-11 tended to be upregulated after A. hydrophila challenge, and G3 in different organs was significantly higher than that in other supplemented groups and G0. The results of this study show that an appropriate amount of mixed antimicrobial peptides can improve the growth performance and antioxidant and immune capabilities of Pengze crucian carp and can also play a positive role in the treatment of A. hydrophila infection.

Synergistic and On-Demand Release of Ag-AMPs Loaded on Porous Silicon Nanocarriers for Antibacteria and Wound Healing.

Due to the abuse of antibiotics, antimicrobial resistance is rapidly emerging and becoming a major global risk for public health. Thus, there is an urgent need for reducing the use of antibiotics, finding novel treatment approaches, and developing controllable release systems. In this work, a dual synergistic antibacterial platform with on-demand release ability based on silver nanoparticles (AgNPs) and antimicrobial peptide (AMP) coloaded porous silicon (PSi) was developed. The combination of AgNPs and AMPs (Tet-213, KRWWKWWRRC) exhibited an excellent synergistic antibacterial effect. As a carrier, porous silicon can efficiently load AgNPs and AMP under mild conditions and give the platform an on-demand release ability and a synergistic release effect. The AgNPs and AMP coloaded porous silicon microparticles (AgNPs-AMP@PSiMPs) exhibited an acid pH and reactive oxygen species (ROS)-stimulated release of silver ions (Ag+) and AMPs under bacterial infection conditions because of oxidation and desorption effects. Moreover, the release of the bactericide could be promoted by each other due to the interplay between AgNPs and Tet-213. In vitro antibacterial tests demonstrated that AgNPs-AMP@PSiMPs inherited the intrinsic properties and synergistic antibacterial efficiency of both bactericides. In addition, wound dressing loaded with AgNPs-AMP@PSiMPs showed outstanding in vivo bacteria-killing activity, accelerating wound-healing, and low biotoxicity in aStaphylococcus aureus-infected rat wound model. The present work demonstrated that PSiMPS might be an efficient platform for loading the antibiotic-free bactericide, which could synergistically and on-demand release to fight wound infection and promote wound healing.

The role of microbial infection in the pathogenesis of Alzheimer's disease and the opportunity for protection by anti-microbial peptides.

Alzheimer's disease (AD) is the most common cause of dementia. Its pathology is primarily characterized by extracellular deposits of amyloid ß peptide and intracellular neurofibrillary tangles. Current rationales to explain the pathogenesis of AD include amyloid cascade, inflammation, infection defense and anti-microbial protection hypotheses. This review focuses on recent advances in the infection hypothesis, in particular on those pathogenic microbes that act systemically, via periodontal and gastro-intestinal infection routes. It is proposed that the evidence convincingly supports that pathogenic microbial infection is associated with, and is likely a causative trigger for, AD pathology. Microbes can drive AD pathology by two main pathways: either by directly infecting the brain and stimulating amyloid-mediated defence (causative trigger) or indirectly, by stimulating the pro-inflammatory effects of infection. In this context, it follows that anti-microbial/anti-infection therapies could be effective for regulating the pathology and symptoms of AD, depending on the stage of disease. As long-term administration of traditional antibiotic therapy is not recommended, alternative antibiotic agents such as anti-microbial peptides (AMPs), could be preferred for intervention and disease management of AD.

Challenge to overcome current limitations of cell-penetrating peptides.

The penetration of biological membranes is a prime obstacle for the delivery of pharmaceutical drugs. Cell-penetrating peptide (CPP) is an efficient vehicle that can deliver various cargos across the biological membranes. Since the discovery, CPPs have been rigorously studied to unveil the underlying penetrating mechanism as well as to exploit CPPs for various biomedical applications. This review will focus on the various strategies to overcome current limitations regarding stability, selectivity, and efficacy of CPPs.

Studies on antimicrobial peptide-loaded nanomaterial for root caries restorations to inhibit periodontitis related pathogens in periodontitis care.

AIMS: To prepare a novel antimicrobial peptide Nal-P-113 loaded poly (ethylene glycol) combined chitosan nanoparticles (Nal-P-113-PEG-CSNPs) for root caries restorations to control the periodontitis related pathogens in periodontitis care. METHODS: Nanoparticles were prepared by simple polymerisation method and characterised using effective analytical methods (TEM, UV, etc.). The antimicrobial activity and biofilm formation of Nal-P-113-PEG-CSNPs was tested against periodontal bacterial pathogens by different in vitro methods. RESULTS: The size of Nal-P-113 loaded PEG-Chitosn nanoparticles was 216.2 ± 1.6 nm. The drug encapsulation efficiency (%EE (w/w) of Nal-P-113-PEG-CSNPs was found to be 89.33 ± 1.67% (w/w). The antimicrobial examination showed that prepared NPs have effectively inhibited the growth of Fusobacterium nucleatum, Streptococcus gordonii, and Porphyromonas gingivalis with the MIC of 23 µg/mL, 6 µg/mL and 31 µg/mL, respectively. CONCLUSIONS: The prepared antimicrobial peptide-loaded PEG-CSNPs provide excellent in vitro efficiency but, further studies are necessary to confirm its therapeutic efficacy on periodontitis care.

Antibacterial activity of antimicrobial peptide-conjugated nanofibrous membranes.

Antimicrobial peptides (AMPs) are considered as novel potential alternatives to antibiotics against increasing number of multi drug resistant (MDR) pathogens. Although AMPs have shown strong antimicrobial activity against gram-negative or gram-positive microorganisms, AMP conjugated biomaterials that are effective against MDR microorganisms are yet to be developed. Herein, the potential use of (RWRWRWRW)-NH2 (AMP-1) and KRFRIRVRV-NH2 (AMP-2) peptide conjugated electrospun polylactic-co-glycolic-acid (PLGA) nanofibers (NFs) fabricated and their antimicrobial effect by themselves and in their dual combination (1:1) were evaluated on P. aeruginosa and methicillin-resistant S. aureus (MRSA). Those AMP conjugated NFs did not inhibit proliferation of keratinocytes. These results suggest that AMP conjugated NF, which has multiple biological activities, would be a promising candidate as a wound dressing material.

Effects of compound antimicrobial peptides on the growth performance, antioxidant and immune responses and disease resistance of grass carp (Ctenopharyngodon idellus).

In this study, the compound antimicrobial peptides was added to the basic diet to probe its effects on grass carp for 8 weeks. There were 6 groups in our study, including M0 (0 mg/kg) and 5 additional groups: M1 (100 mg/kg), M2 (200 mg/kg), M3 (400 mg/kg), M4 (800 mg/kg) and M5 (1600 mg/kg). The results are as follows: (1) The grass carp's FBW, WGR and SGR in M4 were significantly increased than M0 (p < 0.05). (2) In the hepatopancreas, the amylase activity of M2, M3 and M5 were significant up-regulation than other groups (p < 0.05). (3) The activity of T-AOC in the M3 and M4 was significantly higher compared to the other groups in grass carp hepatopancreas, while MDA was significantly reduced compared to the control group (p < 0.05). The activity of SOD in M5 was significantly increased than the other groups (p < 0.05). (4) The expression of MnSOD (except head kidney), IL8 and TNF-α and IL-1ß (except hepatopancreas) were significantly increased (p < 0.05), while TGF-ß and were significantly reduced in the hepatopancreas, spleen and head kidney at M3 (p < 0.05), and IL10 were significantly decreased in the hepatopancreas at M3 (p < 0.05). In addition, expression of IL8 and TNF-α were significant down-regulation, whereas TGF-ß (expect head kidney) were significant up-regulation in hepatopancreas, spleen and head kidney in M3 after challenge with A. hydrophila. The expression of IL-1ß in spleen and head kidney were also down-regulated, whereas IL10 were significantly up-regulated in the hepatopancreas at M3 after challenge with A. hydrophila (p < 0.05). The results of this study showed that grass carp fed a diet supplemented compound antimicrobial peptides was added with 400-800 mg/kg, which could improve the growth performance, antioxidant and digestive capabilities, and could also enhance the expression of immune-related genes and control to A. hydrophila.

Antimicrobial peptides: a promising strategy for lung cancer drug discovery?

INTRODUCTION: Antimicrobial peptides (AMPs), also called host defense peptides (HDPs), are identified in almost any form of life, which play an important role in innate immune systems. They have a broad spectrum of antifungal, antiviral, antibacterial, and anticancer activities. Lung cancer remains the leading cause of global cancer-related death. Unfortunately, lung cancer chemotherapy is accompanied by serious side effects, nonspecific toxicity, and multidrug resistance. Hence, to overcome these drawbacks, anticancer peptides (ACPs) derived from AMPs may represent a potential promising synergistic treatment strategy for lung cancer. AREAS COVERED: In this review, the authors provide the recent advancements in the use of AMPs for the treatment of lung cancer. Furthermore, the anti-lung cancer modes of action of these peptides have been fully reviewed. Importantly, various strategies for increasing the efficiency and safety of AMPs have been discussed. EXPERT OPINION: The combination of AMPs and other cancer treatment approaches such as chemotherapy, nanoparticle-based delivery systems, and photodynamic therapy can be used as a promising revolutionary strategy for the treatment of lung cancer. The most significant limitations of this strategy that need to be focused on are low efficiency and off-target events.

Towards Robust Delivery of Antimicrobial Peptides to Combat Bacterial Resistance.

Antimicrobial peptides (AMPs), otherwise known as host defence peptides (HDPs), are naturally occurring biomolecules expressed by a large array of species across the phylogenetic kingdoms. They have great potential to combat microbial infections by directly killing or inhibiting bacterial activity and/or by modulating the immune response of the host. Due to their multimodal properties, broad spectrum activity, and minimal resistance generation, these peptides have emerged as a promising response to the rapidly concerning problem of multidrug resistance (MDR). However, their therapeutic efficacy is limited by a number of factors, including rapid degradation, systemic toxicity, and low bioavailability. As such, many strategies have been developed to mitigate these limitations, such as peptide modification and delivery vehicle conjugation/encapsulation. Oftentimes, however, particularly in the case of the latter, this can hinder the activity of the parent AMP. Here, we review current delivery strategies used for AMP formulation, focusing on methodologies utilized for targeted infection site release of AMPs. This specificity unites the improved biocompatibility of the delivery vehicle with the unhindered activity of the free AMP, providing a promising means to effectively translate AMP therapy into clinical practice.

Preparation of aminated fish scale collagen and oxidized sodium alginate hybrid hydrogel for enhanced full-thickness wound healing.

Acute full-thickness wounds require a more extended healing period, thus increasing the risk of infection. Severe infection frequently resulted in wound ulceration, necrosis, and even life-threatening complications. Here, a hybrid hydrogel comprising aminated collagen (AC), oxidized sodium alginate (OSA), and antimicrobial peptides (polymyxin B sulfate and bacitracin) was developed to enhance full-thickness wound healing. The AC with low immunogenicity and high biocompatibility was made from marine fish scales, which are eco-friendly, low-cost, and sustainable. The cross-linked hydrogel was formed by a Schiff base reaction without any catalysts and additional procedures. As expected, the presented hybrid hydrogel can effectively against E. coli and S. aureus, as well as promote cell growth and angiogenesis in vitro. In addition, the hydrogel can promote full-thickness wound healing in a rat model through accelerating reepithelialization, collagen deposition, and angiogenesis. Our work demonstrated that the hybrid hydrogel has promising applications in the field of wound healing, which would prompt the utilization of marine fish resources during food processing.

Acylation of the antimicrobial peptide CAMEL for cancer gene therapy.

Obtaining ideal gene delivery vectors is still a major goal in cancer gene therapy. CAMEL, a short hybrid antimicrobial peptide, can kill cancer cells by membrane lysis. In this study, we constructed a series of non-viral vectors by attaching fatty acids with different chain lengths to the N-terminus of CAMEL. Our results showed that the cellular uptake and transfection efficiency of acyl-CAMEL started to significantly increase from a chain length of 12 carbons. C18-CAMEL was screened for gene delivery because it had the highest transfection efficiency. Surprisingly, C18-CAMEL/plasmid complexes displayed strong endosomal escape activity after entering cells via endocytosis. Importantly, C18-CAMEL could deliver p53 plasmids to cancer cells and significantly inhibited cell proliferation by the expression of p53. In addition, the C18-CAMEL/p53 plasmid complexes and the MDM2 inhibitor nutlin-3a showed significantly synergistic anticancer activity against MCF-7 cells expressing wild-type p53. Conclusively, our study demonstrated that conjugation of stearic acid to antimicrobial peptides is a simple and successful approach for constructing efficient and economical non-viral vectors for cancer gene therapy.

Potentiation of Antibiotics by a Novel Antimicrobial Peptide against Shiga Toxin Producing E. coli O157:H7.

Infection with Shiga toxin-producing Escherichia coli (STEC) results in hemorrhagic colitis and can lead to life-threatening sequelae including hemolytic uremic syndrome (HUS). Conventional treatment is intravenous fluid volume expansion. Antibiotic treatment is contraindicated, due in part to the elevated risk of HUS related to increased Shiga toxin (Stx) release associated with some antibiotics. Given the lack of effective strategies and the increasing number of STEC outbreaks, new treatment approaches are critically needed. In this study, we used an antimicrobial peptide wrwycr, previously shown to enhance STEC killing without increasing Stx production, in combination with antibiotic treatments. Checkerboard and time-kill assays were used to assess peptide wrwycr-antibiotic combinations for synergistic STEC killing. Cytotoxicity and real-time PCR were used to evaluate Stx production and stx expression, respectively, associated with these combinations. The synergistic combinations that showed rapid killing, no growth recovery and minimal Stx production were peptide wrwycr-kanamycin/gentamicin. Transmission electron microscopy revealed striking differences in bacterial cell morphology associated with various treatments. This study provides proof of principle for the design of an antibiotic-peptide wrwycr combination effective in killing STEC without enhancing release of Shiga toxins. It also offers a strategy for the repurposing of antibiotics for treatment of STEC infection.

Hyaluronic acid and antimicrobial peptide-modified gold/silver hybrid nanocages to combat bacterial multidrug resistance.

Bacterial multidrug resistance is a major challenge for the treatment of infection. In this study, a gold-silver hybrid nanocage (Au/Ag NCs) is designed to conjugated with an antimicrobial peptide (AP) and hyaluronic acid (HA) via Au-S bond and electrostatic adsorption respectively. HA-P(Au/Ag) shows a small size (128 nm), a high efficiency of photothermal conversion, and a good stability. Under near-infrared (NIR) irradiation, HA-P(Au/Ag) effectively kills multidrug resistant bacteria-Acinetobacter baumannii (MDR-AB) by disrupting their inner and outer membrane. A pneumonia model caused by MDR-AB is established in mice. HA-P(Au/Ag) treatment reduces the number of bacterial colonies and inflammation in lung tissues and restores the structure of pulmonary alveoli. HA-P(Au/Ag)/NIR treatment increases the survival rate of pneumonia mice to 100%. Safety evaluation demonstrates that HA-P(Au/Ag) causes little cytotoxicity and hemolysis, and shows neglectable impact to the key indicators of kidney and liver function. To conclude, HA-P(Au/Ag) is a highly efficient and safe strategy that is promising to combat MDR-AB caused infection in vitro and in vivo.

Anti-bacterial activity of inorganic nanomaterials and their antimicrobial peptide conjugates against resistant and non-resistant pathogens.

This review details the antimicrobial applications of inorganic nanomaterials of mostly metallic form, and the augmentation of activity by surface conjugation of peptide ligands. The review is subdivided into three main sections, of which the first describes the antimicrobial activity of inorganic nanomaterials against gram-positive, gram-negative and multidrug-resistant bacterial strains. The second section highlights the range of antimicrobial peptides and the drug resistance strategies employed by bacterial species to counter lethality. The final part discusses the role of antimicrobial peptide-decorated inorganic nanomaterials in the fight against bacterial strains that show resistance. General strategies for the preparation of antimicrobial peptides and their conjugation to nanomaterials are discussed, emphasizing the use of elemental and metallic oxide nanomaterials. Importantly, the permeation of antimicrobial peptides through the bacterial membrane is shown to aid the delivery of nanomaterials into bacterial cells. By judicious use of targeting ligands, the nanomaterial becomes able to differentiate between bacterial and mammalian cells and, thus, reduce side effects. Moreover, peptide conjugation to the surface of a nanomaterial will alter surface chemistry in ways that lead to reduction in toxicity and improvements in biocompatibility.

Vaccine Potential of a Recombinant Bivalent Fusion Protein LcrV-HSP70 Against Plague and Yersiniosis.

To counteract the deadly pathogens, i.e., Y. pestis, Y. enetrocolitica, and Y. pseudotuberculosis, we prepared a recombinant DNA construct lcrV-hsp70 encoding the bivalent fusion protein LcrV-HSP70. The lcrV gene of Y. pestis and hsp70 domain II DNA fragment of M. tuberculosis were amplified by PCR. The lcrV amplicon was first ligated in the pET vector using NcoI and BamHI restriction sites. Just downstream to the lcrV gene, the hsp70 domain II was ligated using BamHI and Hind III restriction sites. The in-frame and the orientation of cloned lcrV-hsp70 were checked by restriction analysis and nucleotide sequencing. The recombinant bivalent fusion protein LcrV-HSP70 was expressed in E. coli and purified by affinity chromatography. The vaccine potential of LcrV-HSP70 fusion protein was evaluated in formulation with alum. BALB/c mice were vaccinated, and the humoral and cellular immune responses were studied. The fusion protein LcrV-HSP70 induced a strong and significant humoral immune response in comparison to control animals. We also observed a significant difference in the expression levels of IFN-γ and TNF-α in LcrV-HSP70-immunized mice in comparison to control, HSP70, and LcrV groups. To test the protective efficacy of the LcrV-HSP70 fusion protein against plague and Yersiniosis, the vaccinated mice were challenged with Y. pestis, Y. enterocolitica, and Y. pseudotuberculosis separately. The bivalent fusion protein LcrV-HSP70 imparted 100% protection against the plague. In the case of Yersiniosis, on day 2 post challenge, there was a significant reduction in the number of CFU of Y. enterocolitica and Y. pseudotuberculosis in the blood (CFU/ml) and the spleen (CFU/g) of vaccinated animals in comparison to the LcrV, HSP70, and control group animals.

The insect antimicrobial peptide cecropin A disrupts uropathogenic Escherichia coli biofilms.

Current antibiotics cannot eradicate uropathogenic Escherichia coli (UPEC) biofilms, leading to recurrent urinary tract infections. Here, we show that the insect antimicrobial peptide cecropin A (CecA) can destroy planktonic and sessile biofilm-forming UPEC cells, either alone or when combined with the antibiotic nalidixic acid (NAL), synergistically clearing infection in vivo without off-target cytotoxicity. The multi-target mechanism of action involves outer membrane permeabilization followed by biofilm disruption triggered by the inhibition of efflux pump activity and interactions with extracellular and intracellular nucleic acids. These diverse targets ensure that resistance to the CecA + NAL combination emerges slowly. The antimicrobial mechanisms of CecA, thus, extend beyond pore-forming activity to include an unanticipated biofilm-eradication process, offering an alternative approach to combat antibiotic-resistant UPEC infections.

On-demand storage and release of antimicrobial peptides using Pandora's box-like nanotubes gated with a bacterial infection-responsive polymer.

Background: Localized delivery of antimicrobial agents such as antimicrobial peptides (AMPs) by a biomaterial should be on-demand. Namely, AMPs should be latent and biocompatible in the absence of bacterial infection, but released in an amount enough to kill bacteria immediately in response to bacterial infection. Methods: To achieve the unmet goal of such on-demand delivery, here we turned a titanium implant with titania nanotubes (Ti-NTs) into a Pandora's box. The box was loaded with AMPs (HHC36 peptides, with a sequence of KRWWKWWRR) inside the nanotubes and "closed" (surface-modified) with a pH-responsive molecular gate, poly(methacrylic acid) (PMAA), which swelled under normal physiological conditions (pH 7.4) but collapsed under bacterial infection (pH ≤ 6.0). Thus, the PMAA-gated Ti-NTs behaved just like a Pandora's box. The box retarded the burst release of AMPs under physiological conditions because the gate swelled to block the nanotubes opening. However, it was opened to release AMPs to kill bacteria immediately when bacterial infection occurred to lowering the pH (and thus made the gate collapse). Results: We demonstrated such smart excellent bactericidal activity against a panel of four clinically important bacteria, including Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus. In addition, this box was biocompatible and could promote the osteogenic differentiation of human mesenchymal stem cells. Both in vitro and in vivo studies confirmed the smart "on-demand" bactericidal activity of the Pandora's box. The molecularly gated Pandora's box design represents a new strategy in smart drug delivery.

Metal Binding Antimicrobial Peptides in Nanoparticle Bio-functionalization: New Heights in Drug Delivery and Therapy.

Peptides are considered very important due to the diversity expressed through their amino acid sequence, structure variation, large spectrum, and their essential role in biological systems. Antimicrobial peptides (AMPs) emerged as a potent tool in therapy owing to their antimicrobial properties but also their ability to trespass the membranes, specificity, and low toxicity. They comprise a variety of peptides from which specific amino acid-rich peptides are of interest to the current review due to their features in metal interaction and cell penetration. Histidine-rich peptides such as Histatins belong to the metal binding salivary residing peptides with efficient antibacterial, antifungal, and wound-healing activities. Furthermore, their ability to activate in acidic environment attracted the attention to their potential in therapy. The current review covers the current knowledge about AMPs and critically assess the potential of associating with metal ions both structurally and functionally. This review provides interesting hints for the advantages provided by AMPs and metal ions in biomedicine, making use of their direct properties in brain diseases therapy or in the creation of new bio-functionalized nanoparticles for cancer diagnosis and treatment.