Centre for Children's Burns and Trauma Research, The University of Queensland, Department of Paediatrics and Child Health and Queensland Children's Medical Research Institute, Level 4, Foundation Building, Royal Children's Hospital, Brisbane, QLD 4029, Australia.
Burns. 2011 Sep;37(6):994-1000. doi: 10.1016/j.burns.2011.03.017. Epub 2011 Apr 22.
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).
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.
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.
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.
我们开发了一种方法来测试我们烧伤中心使用的伤口敷料、软膏、乳膏和凝胶的细胞毒性，将它们放置在多孔 Nunc™ 聚碳酸酯细胞培养插入物上，与单层细胞（HaCaTs 和原代人角质形成细胞）孵育。
我们采用两种不同的方法来确定对细胞的相对毒性。（1）光可视化：将敷料或化合物放置在插入物的膜上，然后将其放置在单层组织培养板上。24 小时后，用甲苯胺蓝染色存活的贴壁细胞，并拍摄平板照片。用缓冲液冲洗掉的非贴壁死亡细胞的无细胞区域被测量为平板总面积的百分比。（2）存活细胞的细胞计数：用测试材料孵育 24 小时后，用胰蛋白酶分离剩余的细胞，离心并在含台盼蓝的血细胞计数器中计数，台盼蓝可区分活细胞和死细胞。
测试了 17 种产品。Melolite™、White soft Paraffin™ 和 Chlorsig1% Ointment 是最具细胞毒性的产品。Jelonet™、Mepitel(®)、PolyMem(®)、DuoDerm(®) 和 Xeroform™ 显示出一定的细胞毒性。最具细胞毒性的产品包括含有银或洗必泰的产品，以及含有防腐剂氯甲酚的保湿剂 Paraffin Cream™。
这种体外细胞培养插入物方法允许在无直接细胞接触的情况下测试试剂。它易于操作且快速，应该有助于临床医生确定各种敷料的相对细胞毒性以及每个个体伤口的最佳敷料。
