A Topical Wound Disinfectant (Ethacridine Lactate) Differentially Affects the Production of Immunoregulatory Cytokines in
- 0 Comments
- 9643 reads
A ppropriate wound management is a major issue in hospital care involving almost all fields of medical care. Despite the prophylactic use of a variety of clinically efficient antibiotics in many surgical procedures, the concomitant use of local antiseptics still represents a common and useful therapeutic measure to control wound infection. Particularly for immunocompromised patients, topical antiseptics have to be safe, ie, they must not impair the physiological wound healing process. However, recent publications1–4 reported various interactions of commonly used local antiseptics using cell culture systems or animal models, respectively. These results suggest that local antiseptics profoundly affect not only the viability but also key functions of skin cell types, such as keratinocytes or fibroblasts, or even effector activities of peripheral leukocytes. The latter are inevitably involved in wound healing by producing a variety of regulatory cytokines.5 Nevertheless, reports showing a direct influence of topical antimicrobials on cells of the immune system remain scarce. The aim of the present study was, therefore, to investigate the effects of ethacridine lactate on granulocytes, monocytes, and T-lymphocytes and to screen for immunopharmacological activities beyond its antibacterial properties. In order to mimic the conditions in vivo for the contact of ethacridine lactate with immune cells, the authors used human whole-blood cultures prepared from 6 healthy volunteer donors. With this system, the capacity of the drug to influence the co-stimulated synthesis of several important mediators known to orchestrate wound healing was investigated. Specifically, the authors assessed the production of interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-12 (IL-12), prostaglandin E2 (PGE2), interferon-gamma (IFNγ), interleukin-5 (IL-5), and elastase. Moreover, it was of interest to examine the effects of ethacridine lactate in situations resembling impaired wound healing, leading to chronic wounds. This was studied by adding hydrocortisone to the whole-blood cultures, followed by an incubation with ethacridine lactate. In this part of the experiments, elastase, TGF-beta 1, IL-6, and MCP-1 were determined.
Volunteers and blood collection. For preparation of the cell cultures, blood of 6 healthy volunteers of both genders was used. Donors with symptoms of acute infection or with known chronic inflammatory diseases were excluded from the study. The same applied for individuals receiving a vaccination, surgery, or medications within the last 14 days before donation. The blood was drawn into a syringe prefilled with sodium-heparin (50 IU/mL of blood, ratiopharm, Ulm, Germany) and distributed into the wells of microculture plates (96-well plates, Greiner, Germany).
Test substances. Ethacridine lactate was supplied by Chinosolfabrik (Seelze, Germany) and diluted to the desired concentrations with Hanks Balanced Salt Solution (HBSS, Biochrom KG, Berlin, Germany). Hydrocortisone was purchased from Sigma-Aldrich (Deisenhofen, Germany).
Whole-blood culture system (WBCS) and stimulation of mediator production. The whole-blood culture was performed according to the method described by Meyaard.6 Briefly, after seeding the diluted blood into 96-well plates, ethacridine lactate was added in 5 different concentrations: 10, 3, 1, 0.3, and 0.1 µg/mL (final concentration in culture). All cultures were done in triplicate. The stimuli to induce the production of the different mediators were added. The type of stimulus used allowed discrimination between the responding cell type in the whole blood.
1. Cooper ML, Boyce ST, Hansbrough JF, Foreman TJ, Frank DH. Cytotoxicity to cultured human keratinocytes of topical antimicrobial agents. J Surg Res. 1990;48(3):190–195.
2. Cooper ML, Laxer JA, Hansbrough JF. The cytotoxic effects of commonly used topical antimicrobial agents on human fibroblasts and keratinocytes. J Trauma. 1991;31(6):775–782.
3. Kjolseth D, Frank JM, Barker JH, et al. Comparison of the effects of commonly used wound agents on epithelialization and neovascularization. J Am Coll Surg. 1994;179(3):305–312.
4. Bennett LL, Rosenblum RS, Perlov C, Davidson JM, Barton RM, Nanney LB. An in vivo comparison of topical agents on wound repair. Plast Reconstr Surg. 2001;108(3):675–687.
5. Thomson PD. Immunology, microbiology, and the recalcitrant wound. Ostomy Wound Manage. 2000;46(1A Suppl):77S–82S.
6. Meyaard L, Hovenkamp E, Pakker N, van der Pouw Kraan TC, Miedema F. Interleukin-12 (IL-12) production in whole blood cultures from human immunodeficiency virus-infected individuals studied in relation to IL-10 and prostaglandin E2 production. Blood. 1997;89(2):570–576.
7. Essa S, Raghupathy R, Pacsa AS, El-Shazly A, Said T, Azizieh F. Th1-type cytokines production is decreased in kidney transplant recipients with active cytomegalovirus infection. J Med Virol. 2000;60(2):223–229.
8. Engelhardt E, Toksoy A, Goebeler M, Debus S, Brocker EB, Gillitzer R. Chemokines IL-8, GROalpha, MCP-1, IP-10, and Mig are sequentially and differentially expressed during phase-specific infiltration of leukocyte subsets in human wound healing. Am J Pathol. 1998;153(6):1849–1860.
9. Hagedorn M, Hauptmann S, Essinger U, Kaden P, Mittermayer C. [In vitro and in vivo studies of local disinfection and wound healing]. Hautarzt. 1995;46(5):319–324.
10. Hidalgo E, Bartolome R, Dominguez C. Cytotoxicity mechanisms of sodium hypochlorite in cultured human dermal fibroblasts and its bactericidal effectiveness. Chem Biol Interact. 2002;139(3):265–282.
11. Hansbrough JF, Zapata-Sirvent RL, Cooper ML. Effects of topical antimicrobial agents on the human neutrophil respiratory burst. Arch Surg. 1991;126(5):603–608.
12. Zapata-Sirvent RL, Hansbrough JF. Cytotoxicity to human leukocytes by topical antimicrobial agents used for burn care. J Burn Care Rehabil. 1993;14(2 Pt 1):132–140.
13. Witte MB, Barbul A. General principles of wound healing. Surg Clin North Am. 1997;77(3):509–528.
14. Slavin J. The role of cytokines in wound healing. J Pathol. 1996;178(1):5–10.
15. Letterio JJ, Roberts AB. Regulation of immune responses by TGF-beta. Annu Rev Immunol. 1998;16:137–161.
16. Fitzpatrick DR, Bielefeldt-Ohmann H. Transforming growth factor beta in infectious disease: always there for the host and the pathogen. Trends Microbiol. 1999;7(6):232–236.
17. Cordeiro MF. Beyond Mitomycin: TGF-beta and wound healing. Prog Retin Eye Res. 2002;21(1):75–89.
18. Fowler S, Powrie F. Control of immune pathology by IL-10-secreting regulatory T cells. Springer Semin Immunopathol. 1999;21(3):287–294.
19. Pretolani M. Interleukin-10: an anti-inflammatory cytokine with therapeutic potential. Clin Exp Allergy. 1999;29(9):1164–1171.
20. Asadullah K, Docke WD, Sabat RV, Volk HD, Sterry W. The treatment of psoriasis with IL-10: rationale and review of the first clinical trials. Expert Opin Investig Drugs. 2000;9(1):95–102.
21. Rojas IG, Padgett DA, Sheridan JF, Marucha PT. Stress-induced susceptibility to bacterial infection during cutaneous wound healing. Brain Behav Immun. 2002;16(1):74–84.
22. Trinchieri G. Interleukin-12: a cytokine at the interface of inflammation and immunity. Adv Immunol. 1998;70:83–243.
23. van Furth R. Human monocytes and cytokines. Res Immunol. 1998;149(7-8):719–720.
24. Ziesche R, Block LH. Mechanisms of antifibrotic action of interferon gamma-1b in pulmonary fibrosis. Wien Klin Wochenschr. 2000;112(18):785–790.
25. Mateo RB, Reichner JS, Albina JE. Interleukin-6 activity in wounds. Am J Physiol. 1994;266(6 Pt 2):R1840–R1844.
26. Paquet P, Pierard GE. Interleukin-6 and the skin. Int Arch Allergy Immunol. 1996;109(4):308–317.
27. Gallucci RM, Simeonova PP, Matheson JM, et al. Impaired cutaneous wound healing in interleukin-6-deficient and immunosuppressed mice. FASEB J. 2000;14(15):2525–2531.
28. Sugawara T, Gallucci RM, Simeonova PP, Luster MI. Regulation and role of interleukin 6 in wounded human epithelial keratinocytes. Cytokine. 2001;15(6):328–336.
29. Trinchieri G. Immunobiology of interleukin-12. Immunol Res. 1998;17(1-2):269–278.