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OBJECTIVE: There is currently a wide range of cleansing and irrigation solutions available for wounds, many of which contain antimicrobial agents. The aim of this study was to assess the safety of HydroClean Solution (HARTMANN, Germany), a polyhexamethylene biguanide (PHMB)-containing irrigation solution, in a standard cytotoxicity assay, and to assess its effect in a three-dimensional (3D) full-thickness model of human skin. METHOD: A number of commercially available wound cleansing and irrigation solutions, including the PHMB-containing irrigation solution, were tested in a cytotoxicity assay using L929 mouse fibroblasts (ISO 10993-5:2009). The PHMB-containing irrigation solution was then assessed in an in vitro human keratinocyte-fibroblast 3D full-thickness wounded skin model to determine its effect on wound healing over six days. The effect of the PHMB-containing irrigation solution on tissue viability was measured using a lactate dehydrogenase (LDH) assay, and proinflammatory effects were measured using an interleukin-6 (IL-6) production assay. RESULTS: The PHMB-containing irrigation solution was shown to be equivalent to other commercially available cleansing and irrigation solutions when tested in the L929 fibroblast cytotoxicity assay. When assessed in the in vitro 3D human full-thickness wound healing model, the PHMB-containing irrigation solution treatment resulted in no difference in levels of LDH or IL-6 when compared with levels produced in control Dulbecco's phosphate-buffered saline cultures. There was, however, a pronounced tissue thickening of the skin model in the periwound region. CONCLUSION: The experimental data presented in this study support the conclusion that the PHMB-containing irrigation solution has a safety profile similar to other commercially available cleansing and irrigation solutions. Evidence also suggests that the PHMB-containing irrigation solution does not affect tissue viability or proinflammatory cytokine production, as evidenced by LDH levels or the production of IL-6 in a 3D human full-thickness wound healing model. The PHMB-containing irrigation solution stimulated new tissue growth in the periwound region of the skin model.

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OBJECTIVE: Antiseptics are widely used in wound management to prevent or treat wound infections, and have been shown to have antibiofilm efficacy. The objective of this study was to assess the effectiveness of a polyhexamethylene biguanide (PHMB)-containing wound cleansing and irrigation solution on model biofilm of pathogens known to cause wound infections compared with a number of other antimicrobial wound cleansing and irrigation solutions. METHOD: Staphylococcus aureus and Pseudomonas aeruginosa single-species biofilms were cultured using microtitre plate and Centers for Disease Control and Prevention (CDC) biofilm reactor methods. Following a 24-hour incubation period, the biofilms were rinsed to remove planktonic microorganisms and then challenged with wound cleansing and irrigation solutions. Following incubation of the biofilms with a variety of concentrations of the test solutions (50%, 75% or 100%) for 20, 30, 40, 50 or 60 minutes, remaining viable organisms from the treated biofilms were quantified. RESULTS: The six antimicrobial wound cleansing and irrigation solutions used were all effective in eradicating Staphylococcus aureus biofilm bacteria in both test models. However, the results were more variable for the more tolerant Pseudomonas aeruginosa biofilm. Only one of the six solutions (sea salt and oxychlorite/NaOCl-containing solution) was able to eradicate Pseudomonas aeruginosa biofilm using the microtitre plate assay. Of the six solutions, three (a solution containing PHMB and poloxamer 188 surfactant, a solution containing hypochlorous acid (HOCl) and a solution containing NaOCl/HOCl) showed increasing levels of eradication of Pseudomonas aeruginosa biofilm microorganisms with increasing concentration and exposure time. Using the CDC biofilm reactor model, all six cleansing and irrigation solutions, except for the solution containing HOCl, were able to eradicate Pseudomonas aeruginosa biofilms such that no viable microorganisms were recovered. CONCLUSION: This study demonstrated that a PHMB-containing wound cleansing and irrigation solution was as effective as other antimicrobial wound irrigation solutions for antibiofilm efficacy. Together with the low toxicity, good safety profile and absence of any reported acquisition of bacterial resistance to PHMB, the antibiofilm effectiveness data support the alignment of this cleansing and irrigation solution with antimicrobial stewardship (AMS) strategies.

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Bacterial toxins are thought to play a role in delayed wound healing in critically colonised and infected wounds. Endotoxins are released from Gram-negative bacteria when they are lysed by host phagocytic cells during an immune response, or by antimicrobial agents, potentially leading to a detrimental effect on the host tissues. Endotoxins can affect all aspects of the wound healing process, leading to delayed healing and contributing to wound chronicity. Release of endotoxins by bacteria can also have serious systemic effects (for example, septic shock) that can lead to high levels of patient mortality. This review summarises the role and implications on wound healing of bacterial endotoxins, describing the impact of endotoxins on the various phases of the wound healing response. There is a paucity of in vivo/clinical evidence linking endotoxins attributed to a wound (via antibiotic treatment) or their release from infecting bacteria with parameters of delayed wound healing. Future work should investigate if this link is apparent and determine the mechanism(s) by which such detrimental effects occur, offering an opportunity to identify possible treatment pathways. This paper describes the phenomenon of antimicrobial-induced endotoxin release and summarises the use of wound dressings to reduce wound bioburden without inducing microbial death and subsequent release of endotoxins, thus limiting their detrimental effects.

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OBJECTIVE: To assess the efficacy of five silver-containing gelling fibre wound dressings against single-species and multispecies biofilms using internally validated, UKAS-accredited in vitro test models. METHOD: Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans single- and multispecies biofilms were cultured using Centres for Disease Control (CDC) biofilm reactors and colony drip flow reactors (CDFR). Following a 72 hour incubation period, the substrates on which biofilms were grown were rinsed to remove planktonic microorganisms and then challenged with fully hydrated silver-containing gelling fibre wound dressings. Following dressing application for 24 or 72 hours, remaining viable organisms from the treated biofilms were quantified. RESULTS: In single-species in vitro models, all five antimicrobial dressings were effective in eradicating Staphylococcus aureus and Pseudomonas aeruginosa biofilm bacteria. However, only one of the five dressings (Hydrofiber technology with combination antibiofilm/antimicrobial technology) was able to eradicate the more tolerant single-species Candida albicans biofilm. In a more complex and stringent CDFR biofilm model, the hydrofiber dressing with combined antibiofilm/antimicrobial technology was the only dressing that was able to eradicate multispecies biofilms such that no viable organisms were recovered. CONCLUSION: Given the detrimental effects of biofilm on wound healing, stringent in vitro biofilm models are increasingly required to investigate the efficacy of antimicrobial dressings. Using accredited in vitro biofilm models of increasing complexity, differentiation in the performance of dressings with combined antibiofilm/antimicrobial technology against those with antimicrobial properties alone, was demonstrated.

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OBJECTIVE: To assess the in vitro antimicrobial performance of a non-medicated hydro-responsive wound dressing (HRWD) on the sequestration and killing of wound relevant microorganisms found on the World Health Organization (WHO) priority pathogens list. METHODS: Suspensions of Pseudomonas aeruginosa, Acinetobacter baumannii and methicillin-resistant Staphylococcus aureus (MRSA) were placed on petri dishes. Dressings were each placed on top, incubated for 30 minutes and then removed from the inoculated petri dish. The surface of the dressings previously in contact with the bacterial suspensions were placed directly onto a tryptone soy agar (TSA) plate and incubated for 24 hours. Dressings were then removed from the TSA plate and the level of bacterial growth on the plates was assessed. Sequestered microorganism viability was assessed using LIVE/DEAD viability kits and visualisation by epifluorescence. RESULTS: Our results indicated that HRWDs sequester and retain Pseudomonas aeruginosa, Acinetobacter baumannii and MRSA within the dressing. Non-medicated HRWDs containing bound PHMB (polyhexamethylene biguanide, HRWD+PHMB) killed the microorganisms sequestered within the dressing matrix. CONCLUSION: These data suggest that non-medicated HRWD+PHMB is an effective against WHO priority pathogens and promoting goal of antimicrobial stewardship in wound care.

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OBJECTIVE: The aim of this in vitro study was to investigate the transfer of viable Pseudomonas aeruginosa biofilm microorganisms following treatment with debridement tools. METHOD: The level of viable biofilm microorganisms transferred by debridement tools was compared following treatment that reflected the clinical practice of each product. RESULTS: A significant level of microorganism transfer was seen in response to the mechanical debridement tool. Minimal transfer of microorganisms was seen when in vitro-established biofilms were treated with hydroresponsive wound dressing + polyhexamethylene biguanide (HRWD+PHMB, HydroClean plus). Less Pseudomonas aeruginosa was recovered from explants exposed to dressings compared with those exposed to debridement tools suggesting that there was less transfer of bacteria by dressings. CONCLUSION: The reduced transfer of viable microorganisms by HRWD+PHMB may be the result of significant binding and retention of microbes by the superabsorbent polymer within the dressing, together with enhanced sequestered bacterial killing within the dressing by polymer-bound PHMB. The high levels of microbial transfer/transmission seen for debridement tools suggests that, in the clinical setting, a significant level of bacterial spread over the wound surface and/or surrounding skin by these cleansing tools is likely.

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OBJECTIVE: The objective of this study was to evaluate the ability of a non medicated, hydro-responsive wound dressing (HRWD) to effectively aid in the removal of bacteria known to reside (and cause infections) within the wound environment. METHOD: A series of in vitro studies were undertaken using Staphylococcus aureus and Pseudomonas aeruginosa biofilms to evaluate the capabilities of the HRWD to disrupt and disperse biofilms. RESULTS: Biofilms can be broken up and dispersed by HRWD and both Staphylococcus aureus and Pseudomonas aeruginosa numbers can be reduced by a greater than log2 reduction in the presence of HRWD. However, no 'active' agents are released into the wound environment that have an antimicrobial effect. CONCLUSION: Overall, these studies have shown that this dressing acts as an effective debridement tool, and there are other 'physical' antimicrobial mechanisms impacting bacterial residence. These mechanisms include 1) breaking up and dispersal of biofilms so that the resultant planktonic bacteria are absorbed by the dressing and then 2) sequestered and retained (trapped) within its matrix. Additionally, when PHMB (polyhexamethylene biguanide) is bound within the dressing core but is not released into the wound environment there is the added antimicrobial effect resulting from 3) physical contact with this antiseptic component. Reducing the pathogenicity of the bacteria still further is the dressings ability to 4) absorb and sequester the damaging proteases released by pathogenic bacteria.

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Anti-infectives used to treat chronic exuding wounds are diluted by wound exudates, absorbed into dressings, metabolised by proteases and destroyed by pH. In order to mimic such effects of exudates, the efficacy of six topical wound agents was assessed undiluted and at 10% concentrations, including povidone-iodine ointment and a silver-impregnated wound dressing, to remove biofilms of Pseudomonas aeruginosa, multi-species biofilms of Candida albicans and methicillin-resistant Staphylococcus aureus (MRSA) in vitro in a Centers for Disease Control and Prevention (CDC) reactor. Povidone-iodine was also diluted to 3·3% and 33·3% of the commercial concentrations. Viable microorganisms in each preparation were quantified by colony count. No viable P. aeruginosa biofilm material was recovered after 4 and 24 hours of treatment with povidone-iodine ointment at the 100% and 10% concentrations. No C. albicans/MRSA biofilm material was recovered after 4 and 24 hours of treatment with povidone-iodine ointment at the 100% concentration. In general, following dilution, povidone-iodine ointment appeared to exhibit greater biofilm removal than the other agents tested. Further research involving different microorganisms in vitro and in vivo over a longer period of time will help elucidate the full potential of povidone-iodine ointment and liposomal hydrogel.

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The free swell and absorption capacity under compression of six wound dressings that are indicated for moderately to highly exuding wounds was investigated. Measuring in vitro the absorptive capacity and retention under compression is important in terms of clinical efficacy and efficiency. This in vitro comparative study demonstrated that sorbion sachet EXTRA had the highest free swell capacity of the six test dressings and absorbed more than twice the volume (126%) of the test solution than its nearest competitor. When measuring capacity under compression, sorbion sachet EXTRA absorbed 88% more fluid than the nearest competitor.

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Wound research is an evolving science in the equine species. In particular, interest is growing regarding the role that microorganisms play in delaying both acute and chronic wound healing. Equine wounds, particularly lower limb wounds, frequently display delayed healing and infection is commonly the underlying reason. This review will summarize the current research and knowledge surrounding equine wound healing and wound care. Particular focus is placed on the role that microbes play in chronic equine wounds and the significance of associated bacterial biofilms. .