P Kwesiga Maria, Cook Emily, Hannon Jennifer, Wayward Sarah, Gwaltney Caroline, Rao Smitha, C Frost Megan
Department of Biomedical Engineering, Michigan Technological University, Houghton, MI 49931, USA.
Doctoral Program in Physical Therapy, Central Michigan University, Mt. Pleasant, MI 48859, USA.
Med Sci (Basel). 2018 Nov 9;6(4):99. doi: 10.3390/medsci6040099.
Diabetic foot ulcers (DFU) are a major health problem associated with diabetes mellitus. Impaired nitric oxide (NO) production has been shown to be a major contributor to the dysregulation of healing in DFU. The level of impairment is not known primarily due to challenges with measuring NO. Herein, we report the actual level of NO produced by human dermal fibroblasts cultured under normal and high glucose conditions. Fibroblasts produce the extracellular matrix, which facilitate the migration of keratinocytes to close wounds. The results show that NO production was significantly higher in normal glucose compared to high glucose conditions. The real-time NO detected was compared to the nitrite present in the culture media and there was a direct correlation between real-time NO and nitrite in normal glucose conditions. However, real-time NO detection and nitrite measurement did not correlate under high glucose conditions. The inducible nitric oxide synthase (iNOS) enzyme responsible for NO production was upregulated in normal and high glucose conditions and the proliferation rate of fibroblasts was not statistically different in all the treatment groups. Relying only on nitrite to assess NO production is not an accurate determinant of the NO present in the wound bed in pathological states such as diabetes mellitus.
糖尿病足溃疡（DFU）是与糖尿病相关的一个主要健康问题。一氧化氮（NO）生成受损已被证明是DFU愈合失调的主要原因。由于测量NO存在挑战，损伤程度尚不清楚。在此，我们报告在正常和高糖条件下培养的人皮肤成纤维细胞产生的NO的实际水平。成纤维细胞产生细胞外基质，促进角质形成细胞迁移以闭合伤口。结果表明，与高糖条件相比，正常葡萄糖条件下的NO生成显著更高。将检测到的实时NO与培养基中的亚硝酸盐进行比较，在正常葡萄糖条件下，实时NO与亚硝酸盐之间存在直接相关性。然而，在高糖条件下，实时NO检测与亚硝酸盐测量不相关。负责NO生成的诱导型一氧化氮合酶（iNOS）在正常和高糖条件下均上调，且所有治疗组中成纤维细胞的增殖率无统计学差异。仅依靠亚硝酸盐来评估NO生成并不是糖尿病等病理状态下伤口床中NO存在情况的准确决定因素。
