Bactericidal and Cytotoxic Effects of Chloramine-T on Wound Pathogens and Human Fibroblasts In Vitro

Kloth, Luther C.; Berman, Joseph; Laatsch, Linda J.; Kirchner, Phyllis A.

doi:10.1097/01.asw.0000276408.53632.0b

Cited by 12 publications

(16 citation statements)

References 24 publications

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“…Therefore, the antiseptic of choice would be one that provides the utmost, fastest, and most prolonged antimicrobial activity with the least potential for local or systemic toxicity 16 . There are few studies that have evaluated in parallel both antibacterial power and cytotoxicity in vitro 16–19 . The application of a co‐culture system of human cells and bacteria further expands the suggested parallel designs for testing antibacterial power and cytotoxicity.…”

Section: Discussionmentioning

confidence: 99%

HaCaT keratinocytes in co‐culture with Staphylococcus aureus can be protected from bacterial damage by polihexanide

Wiegand

Abel

Ruth

et al. 2009

Wound Repair Regeneration

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Wound healing is compromised by critical colonization and infection with bacteria. Hence, antimicrobial agents are used clinically to decrease the bacterial load and promote wound healing. Polihexanide (PHMB) has been found to be effective against a broad spectrum of micro-organisms and is increasingly utilized in rinsing solutions or in combination with wound dressings because of its good biocompatibility. In the present study, a co-culture of human keratinocytes and Staphylococcus aureus was established to serve as an in vitro model for infected wounds. Incubation of keratinocytes with increasing concentrations of S. aureus led to a dose-dependent decline of cell viability and proliferation. Lactate dehydrogenase release and interleukin-8 liberation were found to be elevated under these conditions. Polihexanide dose-dependently was able to protect keratinocytes from bacterial damage and re-establish normal human cell proliferation in vitro. Furthermore, a dressing consisting of biocellulose derived from Acetobacter xylinum with the addition of polihexanide was adept to safeguard keratinocytes against S. aureus. In conclusion, the co-culture system presented embodies a valuable tool as a model system for infected cells in a non-healing wound. Furthermore, the results obtained support the favorable function of polihexanide in the treatment of infected chronic wounds.

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Section: Discussionmentioning

confidence: 99%

HaCaT keratinocytes in co‐culture with Staphylococcus aureus can be protected from bacterial damage by polihexanide

Wiegand

Abel

Ruth

et al. 2009

Wound Repair Regeneration

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“…The adherence and growth of fibroblasts in the presence of the [CEL + CS] composites were assessed with modifications from Kloth et al 23 Essentially, human fibroblasts (ATCC CRL-2522) were grown in minimal essential medium (MEM) supplemented with 10% FBS and 0.25 mg/mL gentamicin according to ATCC guidelines until at least the 2nd passage. Cells were seeded into the wells containing membrane composites at a concentration of 8 × 10 4 cells/mL per well.…”

Section: Methodsmentioning

confidence: 99%

Chitosan–cellulose composite for wound dressing material. Part 2. Antimicrobial activity, blood absorption ability, and biocompatibility

et al. 2014

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Chitosan (CS), a polysaccharide derived from chitin, the second most abundant polysaccharide, is widely used in the medical world because of its natural and nontoxic properties and its innate ability for antibacterial and hemostasis effects. In this study, the novel composites containing CS and cellulose (CEL) (i.e., [CEL + CS]), which we have previously synthesized using a green and totally recyclable method, were investigated for their antimicrobial activity, absorption of anticoagulated whole blood, anti-inflammatory activity through the reduction of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and the biocompatibility with human fibroblasts. The [CEL + CS] composites were found to inhibit the growth of both Gram positive and negative microorganisms. For examples, the regenerated 100% lyophilized chitosan material was found to reduce growth of Escherichia coli (ATCC 8739 and vancomycin resistant Enterococcus faecalis (ATCC 51299) by 78, 36, and 64%, respectively. The composites are nontoxic to fibroblasts; that is, fibroblasts, which are critical to the formation of connective tissue matrix were found to grow and proliferate in the presence of the composites. They effectively absorb blood, and at the same rate and volume as commercially available wound dressings. The composites, in both air-dried and lyophilized forms, significantly inhibit the production of TNF-α and IL-6 by stimulated macrophages. These results clearly indicate that the biodegradable, biocompatible and nontoxic [CEL + CS] composites, particularly those dried by lyophilizing, can be effectively used as a material in wound dressings.

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“…To address this, some recent studies have tried to implement two-dimensional, monolayer coculture systems of cells and bacteria to test antimicrobial agents that can be added to the culture medium. [27][28][29][30] To date, there is no published method on evaluating antimicrobial dressings in coculture systems. Therefore, we designed a novel 3D coculture system to assess the cytotoxicity and antimicrobial properties of antimicrobial scaffolds/wound dressings in one experiment.…”

Section: Discussionmentioning

confidence: 99%

Skin Tissue Engineering for the Infected Wound Site: Biodegradable PLA Nanofibers and a Novel Approach for Silver Ion Release Evaluated in a 3D Coculture System of Keratinocytes and Staphylococcus aureus

Mohiti-Asli

Pourdeyhimi

Loboa

2014

Tissue Engineering Part C: Methods

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Wound infection presents a challenging and growing problem. With the increased prevalence and growth of multidrug-resistant bacteria, there is a mounting need to reduce and eliminate wound infections using methodologies that limit the ability of bacteria to evolve into further drug-resistant strains. A well-known strategy for combating bacterial infection and preventing wound sepsis is through the delivery of silver ions to the wound site. High surface area silver nanoparticles (AgNPs) allowing extensive silver ion release have therefore been explored in different wound dressings and/or skin substitutes. However, it has been recently shown that AgNPs can penetrate into the stratum corneum of skin or diffuse into the cellular plasma membrane, and may interfere with a variety of cellular mechanisms. The goal of this study was to introduce and evaluate a new type of high surface area metallic silver in the form of highly porous silver microparticles (AgMPs). Polylactic acid (PLA) nanofibers were successfully loaded with either highly porous AgMPs or AgNPs and the antimicrobial efficacy and cytotoxicity of the two silver-based wound dressings were assessed and compared. To better mimic the physiological environment in vivo where both human cells and bacteria are present, a novel coculture system combining human epidermal keratinocytes and Staphylococcus aureus bacteria was designed to simultaneously evaluate human skin cell cytotoxicity with antimicrobial efficacy in a three-dimensional environment. We found that highly porous AgMPs could be successfully incorporated in nanofibrous wound dressings, and exhibited comparable antimicrobial efficacy and cytotoxicity to AgNPs. Further, PLA nanofibers containing highly porous AgMPs exhibited steady silver ion release, at a greater rate of release, than nanofibers containing AgNPs. The replacement of AgNPs with the newly introduced AgMPs overcomes concerns regarding the use of nanoparticles and holds great promise as skin substitutes or wound dressings for infected wound sites.

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Bactericidal and Cytotoxic Effects of Chloramine-T on Wound Pathogens and Human Fibroblasts In Vitro

Abstract: In vitro, chloramine-T at 200 ppm for 5 to 20 minutes was effective against 3 virulent gram-positive bacteria without fibroblast damage. At 300 ppm and 3 and 5 minutes, 30% of fibroblasts were damaged and 95% to 100 % of E coli were inhibited, respectively.

Cited by 12 publications

References 24 publications

HaCaT keratinocytes in co‐culture with Staphylococcus aureus can be protected from bacterial damage by polihexanide

HaCaT keratinocytes in co‐culture with Staphylococcus aureus can be protected from bacterial damage by polihexanide

Chitosan–cellulose composite for wound dressing material. Part 2. Antimicrobial activity, blood absorption ability, and biocompatibility

Skin Tissue Engineering for the Infected Wound Site: Biodegradable PLA Nanofibers and a Novel Approach for Silver Ion Release Evaluated in a 3D Coculture System of Keratinocytes and Staphylococcus aureus

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