Excisional Wound Closure Assessment
Objective: Determine the requirement for inducible nitric oxide synthase (iNOS) in excisional wound closure by comparing wound closure rates in iNOS knockout mice versus wild-type controls
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Materials2
Software1
Not specified • Not specified
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Protocol Steps
Create excisional wounds
Excisional wounds were created in iNOS knockout mice and wild-type control animals
Note: Baseline wound creation for comparison between genotypes
View evidence from paper
“Experiments were carried out to determine the requirement for iNOS in closing excisional wounds”
Measure wound closure in iNOS knockout versus wild-type mice
Wound closure was measured and compared between iNOS knockout mice and wild-type animals
Note: Wound closure was delayed by 31% in iNOS knockout mice compared with wild-type animals
View evidence from paper
“Wound closure was delayed by 31% in iNOS knockout mice compared with wild-type animals”
Administer iNOS inhibitor to wild-type mice
Wild-type mice were given continuous infusion of N6-(iminoethyl)-L-lysine, a partially selective iNOS inhibitor
Note: This treatment produced an identical delay in wound closure as observed in iNOS knockout mice
View evidence from paper
“An identical delay in wound closure was observed in wild-type mice given a continuous infusion of the partially selective iNOS inhibitor N6-(iminoethyl)-L-lysine”
Apply adenoviral iNOS vector to iNOS-deficient mice
A single application of adenoviral vector containing human iNOS cDNA (AdiNOS) was applied at the time of wounding
Note: This treatment completely reversed the delayed wound healing phenotype in iNOS-deficient mice
View evidence from paper
“Delayed wound healing in iNOS-deficient mice was completely reversed by a single application of an adenoviral vector containing human iNOS cDNA (AdiNOS) at the time of wounding”
Measure iNOS mRNA expression by reverse transcription PCR
Reverse transcription PCR was performed to identify iNOS mRNA expression in wild-type mice and confirm human iNOS expression in treated animals
Note: iNOS mRNA expression peaked 4-6 days after wounding in wild-type mice; human iNOS expression was confirmed in AdiNOS-treated animals
View evidence from paper
“Reverse transcription PCR identified iNOS mRNA expression in wild-type mice peaking 4-6 d after wounding, and confirmed expression of human iNOS in the adenoviral vector containing human iNOS cDNA-treated animals”