Antimicrobial effectiveness of wound matrices containing native extracellular matrix with polyhexamethylene biguanide.

A variety of wound matrix materials that are designed to help heal both acute and chronic wounds are currently available. Because wounds often encounter opportunistic microbes that can delay healing, the effectiveness of these materials is often suboptimal, resulting in delayed or compromised wound healing. The importance of reducing and controlling wound microbes is well recognised and there are several antimicrobial options available to address this unmet clinical need. This study compares the antimicrobial and wound healing capabilities, both in vivo and in vitro against methicillin-resistant Staphylococcus aureus (MRSA) USA 300, for the following compounds: Collagen Wound Matrix-Anti Microbial (CWM-AM); Collagen Wound Matrix-Anti Microbial XT (CWM-AM XT); Antimicrobial Hydrofiber Wound Dressing (AHWD); Dermal Scaffold with Silver (DRSAg); Collagen Extracellular Matrix (CEM); Collagen Wound Matrix (CWM); Matrix Wound Dressing with Silver (MWDAg); Cadexomer Iodine Gel (CIG); Triple Antibiotic Ointment (TAO); and Antimicrobial Wound Gel (AWG). For the in vitro zone of inhibition assay, AWG and CIG had the largest diffused areas, followed by CWM-AM and CWM-AM XT. Furthermore, CWM-AM, CWM-AM XT, AWG, and CIG exhibited a persistent antimicrobial activity for up to 10 days after incubation. However, in the cytotoxicity studies performed using human fibroblasts, CWM-AM and CWM-AM XT had no detrimental effects in cell proliferation and viability, while AWG and CIG were cytotoxic and prohibitive for cell proliferation. Treatments were then assessed for microbiology and wound healing efficacy using an in vivo porcine deep reticular dermal wound model. CWM-AM XT displayed the greatest in vivo antimicrobial activity against MRSA USA300 and expedited the reepithelialisation at a faster rate than other treatment groups. This study shows that a novel collagen matrix containing an antimicrobial agent can reduce the bacterial load and support healing.

Novel Cyclic Lipopeptides Fusaricidin Analogs for Treating Wound Infections.

Both acute and chronic cutaneous wounds are often difficult to treat due to the high-risk for bacterial contamination. Once hospitalized, open wounds are at a high-risk for developing hospital-associated infections caused by multi drug-resistant bacteria such as Staphylococcus aureus and Pseudomonas aeruginosa. Treating these infections is challenging, not only because of antibiotic resistance, but also due to the production of biofilms. New treatment strategies are needed that will help in both stimulating the wound healing process, as well as preventing and eliminating bacterial wound infections. Fusaricidins are naturally occurring cyclic lipopeptides with antimicrobial properties that have shown to be effective against a variety of fungi and Gram-positive bacteria, with low toxicity. Continuing with our efforts toward the identification of novel cyclic lipopeptides Fusaricidin analogs, herein we report the synthesis and evaluation of the antimicrobial activity for two novel cyclic lipopeptides (CLP), CLP 2605-4 and CLP 2612-8.1 against methicillin resistant S. aureus and P. aeruginosa, respectively, in in vivo porcine full thickness wound model. Both CLPs were able to reduce bacterial counts by approximately 3 log CFU/g by the last assessment day. Peptide 2612-8.1 slightly enhanced the wound healing, however, wounds treated with peptide 2605-4, have shown higher levels of inflammation and impaired wound healing process. This study highlights the importance of identifying new antimicrobials that can combat bacterial infection while not impeding tissue repair.

Effect of Mechanical Debridement and Irrigation With Hypochlorous Acid Wound Management Solution on Methicillin-resistant Staphylococcus aureus Contamination and Healing Deep Dermal Wounds in a Porcine Model.

BACKGROUND: Wound cleansing is an important component of wound management. PURPOSE: This study was conducted to examine the effect of a wound management solution (WMS) containing hypochlorous acid (HOCl) on methicillin-resistant Staphylococcus aureus (MRSA) and healing when used in conjunction with debridement. METHODS: Nineteen (19) deep reticular dermal wounds (22 mm × 22 mm × 3 mm deep) were created on the paravertebral and thoracic areas of 3 female pigs using a specialized electrokeratome. Wounds were separated by at least 5 cm to 7 cm of unwounded skin and inoculated with MRSA. After 72 hours, all wounds were debrided with a curette and irrigated with either the WMS or sterile saline solution twice per day from day 0 to day 4. Wounds then were irrigated once a day until the completion of the study (day 11). Wound tissue specimens were taken using punch biopsy for microbiological and histological analysis on days 4, 8, and 11 post treatment. Percent of wound epithelialized, epithelial thickness (cell layers µm), white cell infiltrate (1 = absent, 2 = mild, 3 = moderate, 4 = marked, 5 = exuberant), and percent of granulation tissue formation were calculated and assessed. Microbiology and histology results were analyzed for significant differences between treatments and among assessment days using one-way analysis of variance and student t-tests. A P value ≤ .05 was considered significant. RESULTS: The WMS effected a bacterial reduction (P ≤ .05) of more than 2.74 ± 0.43 and 1.03 ± 0.22 Log CFU/g in all assessment days compared with baseline before and after debridement, respectively. Percent epithelialization was significantly different between treatments on day 8, only 78.3% and 67.8% for HOCl and saline, respectively (P ≤ .05). No significant differences between treatments were observed for epithelial thickness or granulation tissue formation. CONCLUSION: The combination of debridement and HOCl wound irrigation can significantly reduce MRSA contamination and facilitate the healing process compared to saline irrigation. Clinical studies are needed to confirm these results.

TGF-ß-mediated enhancement of TH17 cell generation is inhibited by bone morphogenetic protein receptor 1α signaling.

The cytokines of the transforming growth factor-ß (TGF-ß) family promote the growth and differentiation of multiple tissues, but the role of only the founding member, TGF-ß, in regulating the immune responses has been extensively studied. TGF-ß is critical to prevent the spontaneous activation of self-reactive T cells and sustain immune homeostasis. In contrast, in the presence of proinflammatory cytokines, TGF-ß promotes the differentiation of effector T helper 17 (TH17) cells. Abrogating TGF-ß receptor signaling prevents the development of interleukin-17 (IL-17)-secreting cells and protects mice from TH17 cell-mediated autoimmunity. We found that the receptor of another member of TGF-ß family, bone morphogenetic protein receptor 1α (BMPR1α), regulates T helper cell activation. We found that the differentiation of TH17 cells from naive CD4+ T cells was inhibited in the presence of BMPs. Abrogation of BMPR1α signaling during CD4+ T cell activation induced a developmental program that led to the generation of inflammatory effector cells expressing large amounts of IL-17, IFN-γ, and TNF family cytokines and transcription factors defining the TH17 cell lineage. We found that TGF-ß and BMPs cooperated to establish effector cell functions and the cytokine profile of activated CD4+ T cells. Together, our data provide insight into the immunoregulatory function of BMPs.

Coordination of different ligands to copper(II) and cobalt(III) metal centers enhances Zika virus and dengue virus loads in both arthropod cells and human keratinocytes.

Trace elements such as copper and cobalt have been associated with virus-host interactions. However, studies to show the effect of conjugation of copper(II) or cobalt(III) metal centers to thiosemicarbazone ligand(s) derived from either food additives or mosquito repellent such as 2-acetylethiazole or citral, respectively, on Zika virus (ZIKV) or dengue virus (serotype 2; DENV2) infections have not been explored. In this study, we show that four compounds comprising of thiosemicarbazone ligand derived from 2-acetylethiazole viz., (E)-N-ethyl-2-[1-(thiazol-2-yl)ethylidene]hydrazinecarbothioamide (acetylethTSC) (compound 1), a copper(II) complex with acetylethTSC as a ligand (compound 2), a thiosemicarbazone ligand-derived from citral (compound 3) and a cobalt(III) complex with a citral-thiosemicarbazone ligand (compound 4) increased DENV2 and ZIKV replication in both mosquito C6/36 cells and human keratinocytes (HaCaT cells). Treatment of both cell lines with compounds 2 or 4 showed increased dengue viral titers at all three tested doses. Enhanced dengue viral plaque formation was also noted at the tested dose of 100µM, suggesting higher production of infectious viral particles. Treatment with the compounds 2 or 4 enhanced ZIKV and DENV2 RNA levels in HeLa cell line and primary cultures of mouse bone marrow derived dendritic cells. Also, pre- or post treatments with conjugated compounds 2 or 4 showed higher loads of ZIKV or DENV2 envelope (E) protein in HaCaT cells. No changes in loads of E-protein were found in ZIKV-infected C6/36 cells, when compounds were treated after infection. In addition, we tested bis(1,10-phenanthroline)copper(II) chloride ([Cu(phen)2]Cl2, (compound 5) and tris(1,10-phenanthroline)cobalt(III) chloride ([Co(phen)3]Cl3, (compound 6) that also showed enhanced DENV2 loads. Also, we found that copper(II) chloride dehydrate (CuCl2·2H2O) or cobalt(II) chloride hexahydrate (CoCl2·6H2O) alone had no effects as "free" cations. Taken together, these findings suggest that use of Cu(II) or Co(III) conjugation to organic compounds, in insect repellents and/or food additives could enhance DENV2/ZIKV loads in human cells and perhaps induce pathogenesis in infected individuals or individuals pre-exposed to such conjugated complexes. IMPORTANCE: Mosquito-borne diseases are of great concern to the mankind. Use of chemicals/repellents against mosquito bites and transmission of microbes has been the topic of interest for many years. Here, we show that thiosemicarbazone ligand(s) derived from 2-acetylethiazole or citral or 1,10-phenanthroline upon conjugation with copper(II) or cobalt(III) metal centers enhances dengue virus (serotype 2; DENV2) and/or Zika virus (ZIKV) infections in mosquito, mouse and human cells. Enhanced ZIKV/DENV2 capsid mRNA or envelope protein loads were evident in mosquito cells and human keratinocytes, when treated with compounds before/after infections. Also, treatment with copper(II) or cobalt(III) conjugated compounds increased viral titers and number of plaque formations. These studies suggest that conjugation of compounds in repellents/essential oils/natural products/food additives with copper(II) or cobalt(III) metal centers may not be safe, especially in tropical and subtropical places, where several dengue infection cases and deaths are reported annually or in places with increased ZIKV caused microcephaly.