Cytotoxicity testing of burn wound dressings, ointments and creams: A method using polycarbonate cell culture inserts on a cell culture system

• Published in: Burns 2011-09, Vol.37 (6), p.994-1000
• Main Authors: Kempf, Margit, Kimble, Roy M, Cuttle, Leila
• Format: Article
• Language: English
• Subjects: Bandages - adverse effects; Biocompatible Materials - pharmacology; Biological and medical sciences; Burn care dressings; Burns; Cell Survival - drug effects; Cells, Cultured; Critical Care; Cytotoxicity; Humans; Keratinocytes - cytology; Keratinocytes - drug effects; Medical sciences; Ointments - toxicity; Polycarbonate cell culture insert; Polycarboxylate Cement; Tissue culture; Toxicity Tests, Acute; Traumas. Diseases due to physical agents
• ISSN: 0305-4179; 1879-1409
• DOI: 10.1016/j.burns.2011.03.017

Abstract We have developed a method to test the cytotoxicity of wound dressings, ointments, creams and gels used in our Burn Centre, by placing them on a permeable Nunc™ Polycarbonate cell culture insert, incubated with a monolayer of cells (HaCaTs and primary human keratinocytes). Methods We performed two different methods to determine the relative toxicity to cells. (1) Photo visualisation : The dressings or compounds were positioned on the insert's membrane which was placed onto the monolayer tissue culture plate. After 24 h the surviving adherent cells were stained with Toluidine Blue and photos of the plates were taken. The acellular area of non-adherent dead cells which had been washed off with buffer was measured as a percentage of the total area of the plate. (2) Cell count of surviving cells : After 24 h incubation with the test material, the remaining cells were detached with trypsin, spun down and counted in a Haemocytometer with Trypan Blue, which differentiates between live and dead cells. Results Seventeen products were tested. The least cytotoxic products were Melolite™, White soft Paraffin™ and Chlorsig1% Ointment. Some cytotoxicity was shown with Jelonet™, Mepitel® , PolyMem® , DuoDerm® and Xeroform™. The most cytotoxic products included those which contained silver or Chlorhexidine and Paraffin Cream™ a moisturizer which contains the preservative Chlorocresol. Conclusion This in vitro cell culture insert method allows testing of agents without direct cell contact. It is easy and quick to perform, and should help the clinician to determine the relative cytotoxicity of various dressings and the optimal dressing for each individual wound.