Selection of Topical Antimicrobial Agents for Cultured Skin for Burns by Combined Assessment of Cellular Cytotoxicity and Antimicrobial Activity

Cultured epidermal skin has become an adjunctive therapy for treatment of major burn injuries, but its effectiveness is greatly limited due to destruction by microbial contamination. To evaluate candidate drugs for use with cultured skin, a combined cytotoxicity-antimicrobial assay system was developed for determination of toxicity to cultured human keratinocytes and fibroblasts, and to common burn wound organisms (20 bacterial and 4 fungal strains). Candidate agents including Hibiclens (n = 3), amikacin, piperacillin, norfloxacin, and nystatin were tested separately and in combination (n = 6 each) for inhibition of growth of human cells and lytic activity on microorganisms in the wet disc assay. The data showed that: (1) Hibiclens was uniformly toxic to both cultured human cells and microorganisms; (2) norfloxacin had dose-dependent toxicity to human cells and broad effectiveness against microorganisms; and (3) norfloxacin (25 μg/mL) plus nystatin (100 U/mL) had low toxicity to human cells and high toxicity to both Gram-positive and Gram-negative bacteria (20 of 20) and fungi (4 of 4). Selection of topical antimicrobial drugs by these assays may improve effectiveness of cultured skin for burns and may be extended to the control of other surgical wound infections. (Plast. Reconstr. Surg. 92: 493, 1993.)

[1]  D. Greenhalgh,et al.  Skin anatomy and antigen expression after burn wound closure with composite grafts of cultured skin cells and biopolymers. , 1993, Plastic and reconstructive surgery.

[2]  Y. Kuroyanagi,et al.  Evaluation of a synthetic wound dressing capable of releasing silver sulfadiazine. , 1991, The Journal of burn care & rehabilitation.

[3]  D. Herndon,et al.  Topical Bactroban (mupirocin): efficacy in treating burn wounds infected with methicillin-resistant staphylococci. , 1990, The Journal of burn care & rehabilitation.

[4]  I. A. Holder Wet disc testing of mafenide hydrochloride, chlorhexidine gluconate, and triple antibiotic solution against bacteria isolated from burn wounds. , 1990, The Journal of burn care & rehabilitation.

[5]  R. Spence,et al.  Cultured epidermis for the coverage of massive burn wounds. A single center experience. , 1990, Annals of surgery.

[6]  B. Hull,et al.  Coverage of full-thickness burns with bilayered skin equivalents: a preliminary clinical trial. , 1990, Surgery.

[7]  S. Boyce,et al.  Cytotoxicity to cultured human keratinocytes of topical antimicrobial agents. , 1990, The Journal of surgical research.

[8]  R. Dover,et al.  CULTURED COMPOSITE SKIN GRAFTS: BIOLOGICAL SKIN EQUIVALENTS PERMITTING MASSIVE EXPANSION , 1989, The Lancet.

[9]  C. Compton,et al.  Cultured Epithelial Autografts for Giant Congenital Nevi , 1989, Plastic and reconstructive surgery.

[10]  I. A. Holder The wet disc antimicrobial solution assay. An in vitro method to test efficacy of antimicrobial solutions for topical use. , 1989, The Journal of burn care & rehabilitation.

[11]  D. Herndon,et al.  In vitro toxicity of topical antimicrobial agents to human fibroblasts. , 1989, The Journal of surgical research.

[12]  S. Boyce,et al.  Biologic attachment, growth, and differentiation of cultured human epidermal keratinocytes on a graftable collagen and chondroitin-6-sulfate substrate. , 1988, Surgery.

[13]  I. A. Holder,et al.  Norfloxacin and silver-norfloxacin as topical antimicrobial agents: results of in vitro susceptibility testing against bacteria and Candida sp isolated from burn patients. , 1986, The Journal of burn care & rehabilitation.

[14]  C. Compton,et al.  Permanent coverage of large burn wounds with autologous cultured human epithelium. , 1985, The New England journal of medicine.

[15]  S. Boyce,et al.  Cultivation, frozen storage, and clonal growth of normal human epidermal keratinocytes in serum-free media , 1985 .

[16]  W. Lineaweaver,et al.  Cellular and Bacterial Toxicities of Topical Antimicrobials , 1985, Plastic and reconstructive surgery.

[17]  S. Boyce,et al.  Calcium-regulated differentiation of normal human epidermal keratinocytes in chemically defined clonal culture and serum-free serial culture. , 1983, The Journal of investigative dermatology.

[18]  Kevin P. Farrell,et al.  Preliminary experience with cultured epidermal autograft in a community hospital burn unit. , 1992, The Journal of burn care & rehabilitation.