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Establishing Nitrite to Nitrate Ratios in the Clinical Wound Samples of Subjects With and Without Diabetes
Introduction: Diabetes mellitus (DM) has the potential to significantly decrease quality of life due to its complications, including but not limited to diabetic foot ulcers and possible amputation. We hypothesize that the potential use of nitric oxide (NO) to normalize wound healing is clearly suggested. However, information regarding the correct dosage and timing of exogenous NO in the wound bed needs to be determined to bring this potential treatment to clinical use. Problems with understanding the role and use of NO in wound healing stems from the difficulty in directly measuring NO present in the wound site. Previously, other researchers have used values of nitrite and nitrate as a proxy to the level of NO. It has been demonstrated that nitrite and nitrate levels do not correlate with NO production in the wound site. We report the range and variability of nitrite and nitrate levels in subjects with and without diabetes through wound fluid swab samples.
Methods: Subjects were recruited from a wound care clinic. Wound swab samples were taken from 20 subjects with and without diabetes. These samples were analyzed for nitrite and nitrate in the wound bed through a triiodide assay then using chemiluminescence to measure the NO produced with a nitric oxide analyzer.
Results: With both nitrite and nitrate, we found no indicative pattern defining subjects with and without diabetes. Subjects with diabetes had a smaller levels of nitrite and nitrate compared to those without diabetes, but this was not statistically significant (p=0.29 and p=0.26, respectively). The comparison of the ratio of nitrite to nitrate in subjects with and without diabetes was nearing significance (p=0.08).
Discussion: Physiologically, the NO state is very complex, therefore variable factors such as oxidative stress can recycle nitrite and nitrate back into NO. This makes the direct correlation of nitrite and nitrate to NO tenuous at best. Therefore, we conclude nitrite and nitrate values alone in the presence of DM are not accurate predictors of NO in the wound bed.
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