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Addition of epidermal growth factor improves the rate of sulfur mustard wound healing in an in vitro model.

Henemyre-Harris CL, Adkins AL, Chuang AH, Graham JS - Eplasty (2008)

Bottom Line: EGF (1 ng/mL) significantly increased wound fill on all of the days tested (days 6, 9, and 12).KGF did not significantly improve wound healing.These results will be used to develop a dressing that can slowly release EGF on to a debrided wound bed to help speed the healing process.

View Article: PubMed Central - PubMed

Affiliation: Physiology and Immunology Branch, Research Division, US Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD, USA. claudia.henemyre@us.army.mil

ABSTRACT

Objective: Sulfur mustard (SM) causes blisters on the human skin. These blisters delay healing of the skin and make the victims more susceptible to infection. In vitro models have been used for protection studies against SM injury, but study on wound healing after SM exposure has not been explored. The purpose of this study was to test whether the addition of exogenous growth factors could improve the rate of SM wound healing.

Methods: The model consisted of normal human epidermal keratinocytes seeded into 6-well plates, exposed to SM, and wounded (disruption of the cell monolayer) with a sterile wounding instrument. Cells were then stained and images were captured to measure percentage wound fill. Epidermal growth factor (EGF) and keratinocyte growth factor (KGF) were tested in this model.

Results: EGF (1 ng/mL) significantly increased wound fill on all of the days tested (days 6, 9, and 12). KGF did not significantly improve wound healing.

Conclusions: EGF showed promise as a potential therapy for SM-induced wounds. This in vitro model was a valuable tool for screening therapeutics before animal testing. These results will be used to develop a dressing that can slowly release EGF on to a debrided wound bed to help speed the healing process.

No MeSH data available.


Related in: MedlinePlus

Wound fill time-course study for NHEK cells exposed to various concentrations of SM and treated with 1 ng/mL of EGF. Cells were exposed to 0, 5, 10, 25, or 50 μM of SM, treated daily with 0 or 1 ng/mL of EGF, and stained with 0.1% crystal violet at 6 (a), 9 (b), or 12 (c) days after wounding. A significant difference* was observed between the 2 treatment groups, 0 and 1 ng/mL of EGF, on days 6, 9, and 12. However, no significant interactions were observed between SM concentrations and EGF doses, so it cannot be specifically stated that there was a significant difference between EGF doses at a particular SM concentration (ie, 0, 5, 10, 25, or 50 μM of SM). Significant differences between SM concentrations,† regardless of EGF dose, were observed at each day. (a) For day 6, 0 μM of SM† had significantly different wound fill than 5, 10, 25, and 50 μM of SM. In addition, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM. (b and c) For days 9 and 12, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM and 10 μM of SM† had significantly different wound fill than 25 and 50 μM of SM. Data points represent mean values ± SEM of 6 determinations from 2 separate experiments. A 2-factor ANOVA at each staining day was used to compare the SM concentrations and the 2 EGF doses. Statistical significance was defined as P ≤ .05 for all tests. (d) Summary graph of parts (a)–(c).
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Figure 2: Wound fill time-course study for NHEK cells exposed to various concentrations of SM and treated with 1 ng/mL of EGF. Cells were exposed to 0, 5, 10, 25, or 50 μM of SM, treated daily with 0 or 1 ng/mL of EGF, and stained with 0.1% crystal violet at 6 (a), 9 (b), or 12 (c) days after wounding. A significant difference* was observed between the 2 treatment groups, 0 and 1 ng/mL of EGF, on days 6, 9, and 12. However, no significant interactions were observed between SM concentrations and EGF doses, so it cannot be specifically stated that there was a significant difference between EGF doses at a particular SM concentration (ie, 0, 5, 10, 25, or 50 μM of SM). Significant differences between SM concentrations,† regardless of EGF dose, were observed at each day. (a) For day 6, 0 μM of SM† had significantly different wound fill than 5, 10, 25, and 50 μM of SM. In addition, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM. (b and c) For days 9 and 12, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM and 10 μM of SM† had significantly different wound fill than 25 and 50 μM of SM. Data points represent mean values ± SEM of 6 determinations from 2 separate experiments. A 2-factor ANOVA at each staining day was used to compare the SM concentrations and the 2 EGF doses. Statistical significance was defined as P ≤ .05 for all tests. (d) Summary graph of parts (a)–(c).


Addition of epidermal growth factor improves the rate of sulfur mustard wound healing in an in vitro model.

Henemyre-Harris CL, Adkins AL, Chuang AH, Graham JS - Eplasty (2008)

Wound fill time-course study for NHEK cells exposed to various concentrations of SM and treated with 1 ng/mL of EGF. Cells were exposed to 0, 5, 10, 25, or 50 μM of SM, treated daily with 0 or 1 ng/mL of EGF, and stained with 0.1% crystal violet at 6 (a), 9 (b), or 12 (c) days after wounding. A significant difference* was observed between the 2 treatment groups, 0 and 1 ng/mL of EGF, on days 6, 9, and 12. However, no significant interactions were observed between SM concentrations and EGF doses, so it cannot be specifically stated that there was a significant difference between EGF doses at a particular SM concentration (ie, 0, 5, 10, 25, or 50 μM of SM). Significant differences between SM concentrations,† regardless of EGF dose, were observed at each day. (a) For day 6, 0 μM of SM† had significantly different wound fill than 5, 10, 25, and 50 μM of SM. In addition, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM. (b and c) For days 9 and 12, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM and 10 μM of SM† had significantly different wound fill than 25 and 50 μM of SM. Data points represent mean values ± SEM of 6 determinations from 2 separate experiments. A 2-factor ANOVA at each staining day was used to compare the SM concentrations and the 2 EGF doses. Statistical significance was defined as P ≤ .05 for all tests. (d) Summary graph of parts (a)–(c).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2289780&req=5

Figure 2: Wound fill time-course study for NHEK cells exposed to various concentrations of SM and treated with 1 ng/mL of EGF. Cells were exposed to 0, 5, 10, 25, or 50 μM of SM, treated daily with 0 or 1 ng/mL of EGF, and stained with 0.1% crystal violet at 6 (a), 9 (b), or 12 (c) days after wounding. A significant difference* was observed between the 2 treatment groups, 0 and 1 ng/mL of EGF, on days 6, 9, and 12. However, no significant interactions were observed between SM concentrations and EGF doses, so it cannot be specifically stated that there was a significant difference between EGF doses at a particular SM concentration (ie, 0, 5, 10, 25, or 50 μM of SM). Significant differences between SM concentrations,† regardless of EGF dose, were observed at each day. (a) For day 6, 0 μM of SM† had significantly different wound fill than 5, 10, 25, and 50 μM of SM. In addition, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM. (b and c) For days 9 and 12, 5 μM of SM† had significantly different wound fill than 10, 25, and 50 μM of SM and 10 μM of SM† had significantly different wound fill than 25 and 50 μM of SM. Data points represent mean values ± SEM of 6 determinations from 2 separate experiments. A 2-factor ANOVA at each staining day was used to compare the SM concentrations and the 2 EGF doses. Statistical significance was defined as P ≤ .05 for all tests. (d) Summary graph of parts (a)–(c).
Bottom Line: EGF (1 ng/mL) significantly increased wound fill on all of the days tested (days 6, 9, and 12).KGF did not significantly improve wound healing.These results will be used to develop a dressing that can slowly release EGF on to a debrided wound bed to help speed the healing process.

View Article: PubMed Central - PubMed

Affiliation: Physiology and Immunology Branch, Research Division, US Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD, USA. claudia.henemyre@us.army.mil

ABSTRACT

Objective: Sulfur mustard (SM) causes blisters on the human skin. These blisters delay healing of the skin and make the victims more susceptible to infection. In vitro models have been used for protection studies against SM injury, but study on wound healing after SM exposure has not been explored. The purpose of this study was to test whether the addition of exogenous growth factors could improve the rate of SM wound healing.

Methods: The model consisted of normal human epidermal keratinocytes seeded into 6-well plates, exposed to SM, and wounded (disruption of the cell monolayer) with a sterile wounding instrument. Cells were then stained and images were captured to measure percentage wound fill. Epidermal growth factor (EGF) and keratinocyte growth factor (KGF) were tested in this model.

Results: EGF (1 ng/mL) significantly increased wound fill on all of the days tested (days 6, 9, and 12). KGF did not significantly improve wound healing.

Conclusions: EGF showed promise as a potential therapy for SM-induced wounds. This in vitro model was a valuable tool for screening therapeutics before animal testing. These results will be used to develop a dressing that can slowly release EGF on to a debrided wound bed to help speed the healing process.

No MeSH data available.


Related in: MedlinePlus