Dakin solution alters macrophage viability and function



      Macrophages are important in wound defense and healing. Dakin's solution (DS), buffered sodium hypochlorite, has been used since World War I as a topical antimicrobial for wound care. DS has been shown to be toxic to host cells, but effects on immune cells are not well documented.

      Materials and methods

      DS at 0.5%, 0.125%, and ten-fold serial dilutions from 0.25%–0.00025% were evaluated for cellular toxicity on murine macrophages (J774A.1). The effect of DS on macrophage adhesion, phagocytosis, and generation of reactive oxygen species was examined. Macrophage polarization following DS exposure was determined by gene expression using quantitative real-time polymerase chain reaction.


      Concentrations of DS >0.0025% reduced macrophage viability to <5% in exposure times as short as 30 s. Similarly, phagocytosis of Staphylococcus aureus, Pseudomonas aeruginosa, and Aspergillus flavus were significantly reduced at all tested concentrations by macrophages pretreated with DS. H2O2 production was reduced by 8%–38% following treatment with 0.00025%–0.125% DS. Macrophage adherence was significantly increased with >0.0025% DS after 15 min of exposure compared with controls. Quantitative real-time polymerase chain reaction demonstrated that DS exposure resulted in classical macrophage activation, with increased expression of inducible nitric oxide synthase 2, interferon-γ, and interleukin (IL)-1β.


      DS at clinically used concentrations (0.025%–0.25%) was detrimental to macrophage survival and function. For optimal clinical use, understanding the impact of DS on macrophages is important as depletion may result in impaired pathogen clearance and delayed healing. These findings indicate that 0.00025% DS is a safe starting dose; however, optimal use of DS requires further validation with in vivo models.


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        • Rutala W.A.
        • Weber D.J.
        Uses of inorganic hypochlorite (bleach) in health-care facilities.
        Clin Microbiol Rev. 1997; 10: 597
        • Dakin H.D.
        On the use of certain antiseptic substances in the treatment of infected wounds.
        Br Med J. 1915; 2: 318
        • Levine J.M.
        Dakin's solution: past, present, and future.
        Adv Skin Wound Care. 2013; 26: 410
        • Carrel A.
        • Dehelly G.
        The treatment of infected wounds.
        Paul Hoeber, New York1917
        • Bunyan J.
        Treatment of burns and wounds by the envelope method.
        Br Med J. 1941; 2: 1
        • Warkentien T.
        • Rodriguez C.
        • Lloyd B.
        • et al.
        • Infectious Disease Clinical Research Program Trauma Infectious Disease Outcomes Study Group
        Invasive mold infections following combat-related injuries.
        Clin Infect Dis. 2012; 55: 1441
      1. Notes from the field: fatal fungal soft-tissue infections after a tornado—Joplin, Missouri, 2011.
        MMWR Morb Mortal Wkly Rep. 2011; 60: 992
        • Hajdu S.
        • Obradovic A.
        • Presterl E.
        • Vecsei V.
        Invasive mycoses following trauma.
        Injury. 2009; 40: 548
        • Vitrat-Hincky V.
        • Lebeau B.
        • Bozonnet E.
        • et al.
        Severe filamentous fungal infections after widespread tissue damage due to traumatic injury: six cases and review of the literature.
        Scand J Infect Dis. 2009; 41: 491
        • Capoor M.R.
        • Sarabahi S.
        • Tiwari V.K.
        • Narayanan R.P.
        Fungal infections in burns: diagnosis and management.
        Indian J Plast Surg. 2010; 43: S37
        • Fleming A.
        The action of chemical and physiological antiseptics in a septic wound.
        Br J Surg. 1919; 7: 99
        • Hidalgo E.
        • Dominguez C.
        Growth-altering effects of sodium hypochlorite in cultured human dermal fibroblasts.
        Life Sci. 2000; 67: 1331
        • Heggers J.P.
        • Sazy J.A.
        • Stenberg B.D.
        • et al.
        Bactericidal and wound-healing properties of sodium hypochlorite solutions: the 1991 Lindberg Award.
        J Burn Care Rehabil. 1991; 12: 420
        • Lineaweaver W.
        • Howard R.
        • Soucy D.
        • et al.
        Topical antimicrobial toxicity.
        Arch Surg. 1985; 120: 267
        • Wilson J.R.
        • Mills J.G.
        • Prather I.D.
        • Dimitrijevich S.D.
        A toxicity index of skin and wound cleansers used on in vitro fibroblasts and keratinocytes.
        Adv Skin Wound Care. 2005; 18: 373
        • Kozol R.A.
        • Gillies C.
        • Elgebaly S.A.
        Effects of sodium hypochlorite (Dakin's solution) on cells of the wound module.
        Arch Surg. 1988; 123: 420
        • Vick L.R.
        • Propst R.C.
        • Bozeman R.
        • Wysocki A.B.
        Effect of Dakin's solution on components of a dermal equivalent.
        J Surg Res. 2009; 155: 54
        • McKenna P.J.
        • Lehr G.S.
        • Leist P.
        • Welling R.E.
        Antiseptic effectiveness with fibroblast preservation.
        Ann Plast Surg. 1991; 27: 265
        • Cooper M.L.
        • Boyce S.T.
        • Hansbrough J.F.
        • Foreman T.J.
        • Frank D.H.
        Cytotoxicity to cultured human keratinocytes of topical antimicrobial agents.
        J Surg Res. 1990; 48: 190
        • Hansbrough J.F.
        • Zapata-Sirvent R.L.
        • Cooper M.L.
        Effects of topical antimicrobial agents on the human neutrophil respiratory burst.
        Arch Surg. 1991; 126: 603
        • Barsoumian A.
        • Sanchez C.J.
        • Mende K.
        • et al.
        In vitro toxicity and activity of Dakin's solution, mafenide acetate, and amphotericin B on filamentous fungi and human cells.
        J Orthop Trauma. 2013; 27: 428
        • Jiménez-Rubio A.
        • Segura J.J.
        • Llamas R.
        • Jiménez-Planas A.
        • Guerrero J.M.
        • Calvo J.R.
        In vitro study of the effect of sodium hypochlorite and glutaraldehyde on substrate adherence capacity of macrophages.
        J Endod. 1997; 23: 562
        • Segura J.J.
        • Jiménez-Rubio A.
        • Guerrero J.M.
        • Calvo J.R.
        Comparative effects of two endodontic irrigants, chlorhexidine digluconate and sodium hypochlorite, on macrophage adhesion to plastic surfaces.
        J Endod. 1999; 25: 243
        • Mahdavian Delavary B.
        • van der Veer W.M.
        • van Egmond M.
        • Niessen F.B.
        • Beelen R.H.
        Macrophages in skin injury and repair.
        Immunobiology. 2011; 216: 753
        • Ploeger D.T.
        • Hosper N.A.
        • Schipper M.
        • Koerts J.A.
        • de Rond S.
        • Bank R.A.
        Cell plasticity in wound healing: paracrine factors of M1/M2 polarized macrophages influence the phenotypical state of dermal fibroblasts.
        Cell Commun Signal. 2013; 11: 29
        • Deonarine K.
        • Panelli M.C.
        • Stashower M.E.
        • et al.
        Gene expression profiling of cutaneous wound healing.
        J Transl Med. 2007; 5: 11
        • Schaffner A.
        • Douglas H.
        • Braude A.
        Selective protection against conidia by mononuclear and against mycelia by polymorphonuclear phagocytes in resistance to aspergillus. Observations on these two lines of defense in vivo and in vitro with human and mouse phagocytes.
        J Clin Invest. 1982; 69: 617
        • Akpogheneta O.
        • Gil-Lamaignere C.
        • Maloukou A.
        • Roilides E.
        • EUROFUNG Network
        Antifungal activity of human polymorphonuclear and mononuclear phagocytes against non-fumigatus aspergillus species.
        Mycoses. 2003; 46: 77
        • Smeltzer M.S.
        • Thomas J.R.
        • Hickmon S.G.
        • et al.
        Characterization of a rabbit model of staphylococcal osteomyelitis.
        J Orthop Res. 1997; 15: 414
        • Schmidt K.D.
        • Tümmler B.
        • Römling U.
        Comparative genome mapping of pseudomonas aeruginosa PAO with P. aeruginosa C, which belongs to a major clone in cystic fibrosis patients and aquatic habitats.
        J Bacteriol. 1996; 178: 85
        • Qiu H.
        • KuoLee R.
        • Harris G.
        • Van Rooijen N.
        • Patel G.B.
        • Chen W.
        Role of macrophages in early host resistance to respiratory Acinetobacter baumannii infection.
        PLoS One. 2012; 7: e40019
        • Henry-Stanley M.J.
        • Garni R.M.
        • Wells C.L.
        Adaptation of FUN-1 and Calcofluor white stains to assess the ability of viable and nonviable yeast to adhere to and be internalized by cultured mammalian cells.
        J Microbiol Methods. 2004; 59: 289
        • De la Fuente M.
        • Del Rio M.
        • Ferrandez M.D.
        • Hernanz A.
        Modulation of phagocytic function in murine peritoneal macrophages by bombesin, gastrin-releasing peptide and neuromedin C.
        Immunology. 1991; 73: 205
        • Lech M.
        • Gröbmayr R.
        • Weidenbusch M.
        • Anders H.J.
        Tissues use resident dendritic cells and macrophages to maintain homeostasis and to regain homeostasis upon tissue injury: the immunoregulatory role of changing tissue environments.
        Mediators Inflamm. 2012; 2012: 951390
        • Peng D.
        • Huang W.
        • Ai S.
        • Wang S.
        Clinical significance of leukocyte infiltrative response in deep wound of patients with major burns.
        Burns. 2006; 32: 946
        • Asai A.
        • Tsuda Y.
        • Kobayashi M.
        • Hanafusa T.
        • Herndon D.N.
        • Suzuki F.
        Pathogenic role of macrophages in intradermal infection of methicillin-resistant Staphylococcus aureus in thermally injured mice.
        Infect Immun. 2010; 78: 4311
        • Cotter J.L.
        • Fader R.C.
        • Lilley C.
        • Herndon D.N.
        Chemical parameters, antimicrobial activities, and tissue toxicity of 0.1 and 0.5% sodium hypochlorite solutions.
        Antimicrob Agents Chemother. 1985; 28: 118
        • Bennett L.L.
        • Rosenblum R.S.
        • Perlov C.
        • Davidson J.M.
        • Barton R.M.
        • Nanney L.B.
        An in vivo comparison of topical agents on wound repair.
        Plast Reconstr Surg. 2001; 108: 675
        • Lewandowski L.
        • Purcell R.
        • Fleming M.
        • Gordon W.T.
        The use of dilute Dakin's solution for the treatment of angioinvasive fungal infection in the combat wounded: a case series.
        Mil Med. 2013; 178: e503