Is NS-EDTA Effective in Clearing Bacteria From Infected Wounds in a Rat Model?
Authors: Zhu Hongyi , Bao Bingbo , Zheng Xianyou 
Published: June 2018

Abstract

Background
Irrigation is one of the key procedures in open fracture management to eliminate pathogens and prevent infection. Metal ion deprivation could inhibit bacterial adhesins and weaken adhesion to the host tissue. EDTA in solution can competitively bind to a metal ion and thus might be able to inhibit bacterial adhesins.

Questions/purposes
(1) Is normal saline-EDTA toxic to fibroblasts and endothelial cells? (2) In a contaminated wound rat model, does irrigation with normal saline-EDTA solution decrease the risk of positive bacterial cultures and infection when compared with normal saline and soap solutions? (3) In an infected wound rat model, are fewer surgical débridements and irrigations with normal saline-EDTA solution required to obtain culture-free wounds when compared with normal saline and soap controls?

Methods
Normal saline-EDTA solution refers to 1 mmol/L EDTA dissolved in normal saline (pH adjusted to 7.4). Normal saline and soap solutions acted as controls. The toxicity of these solutions to fibroblasts and endothelial cells was assessed in vitro by Annexin V/propidium iodide staining and flow cytometer counting (a well-established method to quantitatively measure the number of dead cells). We established contaminated and infected wound models (bone-exposed or not) with either Staphylococcus aureus or Escherichia coli in rats to investigate the efficacy of normal saline-EDTA solution (n = 30 for the contaminated model and n = 50 for the infected model). For contaminated wounds, the proportion of positive bacterial cultures and infections was compared after irrigation and débridement among the three groups. For infected wounds, we performed irrigation and débridement every 48 hours until the cultures were negative and compared the number of débridements required to achieve a negative culture with survival analysis.

Results
Normal saline-EDTA showed no additional toxicity to fibroblasts and endothelial cells when compared with normal saline (normal saline [97%] versus EDTA [98%] on fibroblasts, p = 0.654; normal saline [97%] versus EDTA [98%] on endothelial cells, p = 0.711). When bone was exposed in the contaminated models, EDTA irrigation resulted in fewer positive bacterial cultures with S aureus (EDTA: 23%, normal saline: 67%, soap: 40%, p = 0.003) and with E coli (EDTA: 27%, normal saline: 57%, soap: 30%, p = 0.032); however, infection risk was only lower with EDTA irrigation (S aureus with EDTA: 10%, normal saline: 33%, soap: 37%, p = 0.039; E coli with EDTA: 3%, normal saline: 27%, soap: 23%, p = 0.038). In the infected wound model, EDTA irrigation resulted in earlier culture-negative wounds (fewer surgical sessions) compared with normal saline and soap solutions (nonbone-exposed wounds infected by S aureus: p = 0.003, infected by E coli: p = 0.001; bone-exposed wounds infected by S aureus: p = 0.012, infected by E coli: p = 0.022).

Conclusions
After in vitro assessment of toxicity and in vivo evaluation of efficacy, we concluded that normal saline-EDTA is superior to normal saline and soap solution in our laboratory models.

Clinical Relevance
The use of normal-saline EDTA as an irrigation solution may reduce the infection rate of wounds. Future studies in large animals and humans might prove our observation in rat models that normal saline-EDTA has an advantage over normal saline as an irrigation solution.