Cytotoxicity and Wound Closure Evaluation in Skin Cell Lines after Treatment with Common Antiseptics for Clinical Use

Ortega-Llamas, Laura; Quiñones-Vico, María I.; García-Valdivia, Marta; Fernández-González, A.; Ubago-Rodríguez, Ana; Torre, Raquel Sanabria-de la; Arias‐Santiago, Salvador

doi:10.3390/cells11091395

Cited by 18 publications

(14 citation statements)

References 40 publications

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“…In both assays, cell migration in the assay of PRM6 was comparable to that of the negative control (cell culture medium) and to that of a commercial CHX-based AMD (Bactigras ® ). This result agrees with in vitro studies that show that the migration capacity of keratinocytes and fibroblasts is not affected when treated with low PHMB concentrations (0.001%) [194]. These two in vitro wound healing assays are not the most representative models of a wound.…”

Section: Biological Evaluation: Wound Healingsupporting

confidence: 88%

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Polyhexanide-Releasing Membranes for Antimicrobial Wound Dressings: A Critical Review

Guiomar

Urbano

2022

Membranes

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The prevalence of chronic, non-healing skin wounds in the general population, most notably diabetic foot ulcers, venous leg ulcers and pressure ulcers, is approximately 2% and is expected to increase, driven mostly by the aging population and the steady rise in obesity and diabetes. Non-healing wounds often become infected, increasing the risk of life-threatening complications, which poses a significant socioeconomic burden. Aiming at the improved management of infected wounds, a variety of wound dressings that incorporate antimicrobials (AMDs), namely polyhexanide (poly(hexamethylene biguanide); PHMB), have been introduced in the wound-care market. However, many wound-care professionals agree that none of these wound dressings show comprehensive or optimal antimicrobial activity. This manuscript summarizes and discusses studies on PHMB-releasing membranes (PRMs) for wound dressings, detailing their preparation, physical properties that are relevant to the context of AMDs, drug loading and release, antibacterial activity, biocompatibility, wound-healing capacity, and clinical trials conducted. Some of these PRMs were able to improve wound healing in in vivo models, with no associated cytotoxicity, but significant differences in study design make it difficult to compare overall efficacies. It is hoped that this review, which includes, whenever available, international standards for testing AMDs, will provide a framework for future studies.

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Section: Biological Evaluation: Wound Healingsupporting

confidence: 88%

“…Depending on the concentration, PHMB may exhibit in vitro cytotoxicity [194]. In PHMB loading by soaking, a dependence of the result of the cytotoxicity evaluation on the conditions employed in the preparation of sample extracts was found in a study with the PRM7 membrane.…”

Section: Prm17mentioning

confidence: 99%

Polyhexanide-Releasing Membranes for Antimicrobial Wound Dressings: A Critical Review

Guiomar

Urbano

2022

Membranes

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“…[ 23 ] Their MSC properties were investigated by the following experiments: Colony‐forming unit assay: hPDLSCs of 4 × 10 3 per well were inoculated in six‐well plates with 2 mL culture medium changed every 2 d. After 3 weeks, the cells were fixed with 4% PFA for 20 min and stained with 1% crystal violet for 1 h. The cell colonies consisting of more than 50 cells were observed using light microscopy (Primo Vert, Carl Zeiss, Jena, Germany). Cell‐doubling time assay: hPDLSCs of 5 × 10 5 per well were inoculated in T175 culture flasks with 20 mL culture medium changed every 2 d. After 3, 6, 9, and 12 d, the cells were collected and counted to produce a cell‐growth curve. Cell viability assay: hPDLSCs of 5 × 10 3 per well were inoculated in 96‐well plates with 0.1 mL culture medium changed every 2 d. After 1, 3, 5, and 7 d, the cell viability was longitudinally assessed by AlamarBlue HS Cell Viability Reagent as described before. [ 56 ] Osteogenic differentiation assay: hPDLSCs of 8 × 10 3 per well were inoculated in 24‐well plates with 0.5 mL culture medium. When the cells reached 80% confluence, the medium was replaced by an osteogenic medium composed of culture medium and 5% StemXVivo human osteogenic supplement.…”

Section: Methodsmentioning

confidence: 99%

“…• Cell viability assay: hPDLSCs of 5 × 10 3 per well were inoculated in 96well plates with 0.1 mL culture medium changed every 2 d. After 1, 3, 5, and 7 d, the cell viability was longitudinally assessed by AlamarBlue HS Cell Viability Reagent as described before. [56] • Osteogenic differentiation assay: hPDLSCs of 8 × 10 3 per well were inoculated in 24-well plates with 0.5 mL culture medium. When the cells reached 80% confluence, the medium was replaced by an osteogenic medium composed of culture medium and 5% StemXVivo human osteogenic supplement.…”

Section: Methodsmentioning

confidence: 99%

Biomimetic Periodontal Ligament Transplantation Activated by Gold Nanoparticles Protects Alveolar Bone

Zong

Holm

Bronckaers

et al. 2023

Adv Healthcare Materials

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Stem cell therapy might be a promising method to stimulate alveolar bone regeneration, which is currently a major clinical challenge. However, its therapeutic features largely depend on pretreatment and transplantation preparation. Herein, a novel biomimetic periodontal ligament transplantation composed of human periodontal ligament stem cells (hPDLSCs) pretreated with gold nanocomplexes (AuNCs) and embedded in a type-I collagen hydrogel scaffold is developed to protect alveolar bone from resorption. AuNCs are readily absorbed by primary hPDLSCs, with limited cytotoxicity, and promote osteogenic differentiation of hPDLSCs effectively in vitro. In addition, the AuNCs-induced hPDLSCs are encapsulated with type-I collagen hydrogel scaffold to mimic their native physiological niche, and then are transplanted into a rat model of alveolar bone resorption. Both micro-computed tomography (micro-CT) and immunohistochemical assays demonstrate that alveolar bone loss is significantly prevented. Furthermore, the underlying therapeutic mechanism is elucidated, in which transplantation-activated osteogenesis is associated with autophagy, which enables bone remodeling and regeneration. This study provides critical insight into the role of PDLSCs in bone homeostasis and proposes an innovative AuNCs-based strategy for stem cell therapy in bone regeneration.

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“…PHMB-based antiseptics introduce low tissue-toxicity, whereas other antiseptics (e.g., PVP-I and octenidine) have significant impact on cell viability 106 (although other studies have shown low cytotoxicity of HOCl to cells 107 ). Laboratory studies have shown that PHMB (or solutions containing PHMB) affect cells in a variety of ways in terms of cytotoxicity: several studies show no or low-level cell cytotoxicity 106,[108][109][110] whilst others show significant levels of toxicity to cells. 111 Kramer al (2019) 104 comparing an extensive number of antiseptics for the antibacterial activity against test organisms and assessing the cytotoxicity on cultured cells found that PHMB was one of the most suitable antimicrobial agents for use wound antisepsis.…”

Section: Broad-spectrum Efficacy Of Phmbmentioning

confidence: 99%

Effectiveness of a polyhexamethylene biguanide-containing wound cleansing solution using experimental biofilm models

Rippon

Rogers

Westgate³

et al. 2023

J Wound Care

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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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Cytotoxicity and Wound Closure Evaluation in Skin Cell Lines after Treatment with Common Antiseptics for Clinical Use

Cited by 18 publications

References 40 publications

Polyhexanide-Releasing Membranes for Antimicrobial Wound Dressings: A Critical Review

Polyhexanide-Releasing Membranes for Antimicrobial Wound Dressings: A Critical Review

Biomimetic Periodontal Ligament Transplantation Activated by Gold Nanoparticles Protects Alveolar Bone

Effectiveness of a polyhexamethylene biguanide-containing wound cleansing solution using experimental biofilm models

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