Kassie Kallestad

Summary
11/11/2015

Outline of: Negative Pressure Wound Therapy Decreases Mortality in a Murine Model of
Burn-Wound Sepsis Involving Pseudomonas aeruginosa Infection
Introduction
Negative Pressure Wound Treatment (NPWT) therapy is a type of wound treatment, using
a vacuum system that continuously pulls moisture out of the wound, there is a foam or gauze
dressing secured by an adhesive to create the negative pressure. (Misaidi, 2012) This method has
been in place since the late 1990s in the U.S. The control of the experiment is a traditional
wound treatment called wet-to-dry (WTD). (Liu, 2014, 2014)This method uses a saline dressing
on the interior and a dry dressing on the exterior, allowing the wound to stay moist while pulling
drainage away from the skin. This method allows enough moisture in the wound to encourage
new cell growth. In some wounds, like burns however, WTD treatment can be a harbor for
bacteria to grow. Since this is a more standard treatment the scientists used this particular method
as the control. (Liu, 2014) Pseudomonas aeruginosa (P. aeruginosa) infection is a very common
infection that can often result in death because of its growing resistance to antibiotics. (Liu,
2014) This study is trying to find out if NPWT could reduce the death rate among mice with
third degree burns infected with P. aeruginosa. Burn victims are at a high risk of getting this
infection because of the shock to their immune system which happens when the body has third
degree burns. (Liu, 2014)According to Liu and colleagues, the research was conducted mainly
because of growing rates of infections and resistance to antibiotics;
The treatment of these infections is confounded by the resistance (both innate and
acquired) of P. aeruginosa to many antimicrobials. It is estimated that 50% of all deaths
caused by burns result from infection, and untreatable forms of P. aeruginosa infection
have unfortunately become more common.(Liu, pg. 1)
This research was done in order to test if NPWT would be more effective in treating and
prolonging the life of an organism that is infected with P. aeruginosa. This type of infection is
very common in humans (especially burn victims) and is constantly adapting making it very
difficult to control via antibiotics. For this reason there are many deaths that occur at the hand of
P. aeruginosa, therefore more research is needed to learn how to effectively treat wounds that are

Kassie Kallestad
Summary
11/11/2015

infected with this bacteria. The researchers hypothesized NPWT would decrease the levels of P.
aeruginosa within a burn wound, prevent sepsis, and reduce the mortality rate of the mice.
Materials and Methods
A total of 342 mice were used in the experiment. Many of the mice were scalded with
third degree burns on their back side. The mice were placed into four groups for this study that
consisted of; a WTD treatment group, a NPWT group, a scald with infection group, and scald
without infection group. Tests were conducted to get bacterial cultures, to analyze cytokine
content, and for mortality analysis. Dividing the mice into four groups allowed them to have a
control group and an experimental group. Throughout the treatment, tissue samples, blood
samples and pictures were taken to test the abundance of the bacteria in the wound site as well as
the immune response of the animal. The tissue samples that were taken were studied to quantify
the white blood cell content and looked for any development or changes in pathologic tissue.
Pathologic tissue is disease prone tissue or tissue that originated via disease. (Liu, 2014) The
blood that was taken was testing the cytokine levels in the blood. Cytokine are types of protein
that are used mainly in immune system communication. (Liu, 2014)
The thermal wounds that were administered to the mice were done using a template to control
the accuracy of the burn size and placement. The bacteria that was applied to the wound site was
cultured with a luminescence property, which allowed the bacteria to be viewed while the mice
were still alive, using the IVIS 100 system. Tissue samples of the skin, liver, kidneys, and lungs
were taken, samples were prepared and dyed, then observed and photographed under 400X
microscope. The cytokine levels were tested by taking a blood sample through the tail vein, then
centrifuged, frozen, thawed, and the cytokines were using enzyme linked immunosorbent assay
kits. The mortality of the mice in each group was recorded and monitored over fourteen days.
Results
The bacteria luminescence portion of the study found that the presence of bacteria in the
WTD versus NPWT were not significant over a seven day period. Except, from twenty-four
hours to seventy-two hours the WTD group had significantly more bacteria than the NPWT. The
WTD and NPWT fought the infection almost equally. The wound biopsy showed that the WTD
treatment group had higher levels of bacteria than the NPWT group had, however, after seven

Kassie Kallestad
Summary
11/11/2015

days the results showed no significant difference in the two test groups bacterial cultures. In the
cytokine analysis, the results for the NPWT group were not elevated very much in any of the
tests. The WTD treatment groups cytokine levels were generally higher than normal levels,
especially on day three of the study. This indicates that the immune systems of the mice in the
WTD group were more threatened that the NPWT group. The mortality analysis resulted in
significant results, after fourteen days 100% of the scald group was still alive, 6.7% of the scald
plus infection group was alive, 33% of the NPWT group survived, and 10% of the WTD
treatment group survived. Supporting the hypothesis, NPWT decreased the mortality rate more
so than WTD treatment against P. aeruginosa infection in burn wounds. In the tissue samples
collected the results were as follows. The wounded skin white blood cell counts were lower in
the NPWT muscle than the WTD treatment group. The lung tissues in the NPWT group was
significantly healthier than the WTD, they did not have thick fluids built up in them and the
white blood cell infiltration was much lower. The result of this showed that NPWT helped reduce
the inflammatory response compared to WTD treatment. The results were what the scientist
hypothesized for the most part. It was surprising that the bioluminescence bacteria testing and
the wound biopsy testing showed no significant data.
Discussion
The results from the study supported the hypothesis. Using NPWT to treat a burn wound
infected by P. aeruginosa was more effective at prolonging life than WTD treatment would in a
similar wound. The study concluded that NPWT discourages the bacteria P. aeruginosa in burn
wound from multiplying quickly, in turn decreasing the chances of death by sepsis. NPWT also
decrease cytokine levels and helped prevent harm to internal organs. The control treatment,
WTD method was the main limitation in the study. It would be optimal if they could use more
than one control to test NPWT against.

Kassie Kallestad
Summary
11/11/2015

Work Cited
Liu, 2014 Y, Zhou Q, Wang Y, Liu, 2014 Z, Dong M, et al. (2014) Negative Pressure Wound
Therapy Decreases Mortality in a Murine Model of Burn-Wound Sepsis Involving Pseudomonas
aeruginosa Infection. PLoS ONE 9(2): e90494. doi:10.1371/journal.pone.0090494
Mirsaidi, Nasrin. Tips and Articles on Device Safety: Negative pressure wound therapy. Food and

Drug Administration, 01/12/2012.

http://www.fda.gov/MedicalDevices/Safety/AlertsandNotices/TipsandArticlesonDeviceSafety/uc
m225038.htm. 10/25/2015.