Investigative Study on Nitric Oxide Production in Human Dermal Fibroblast Cells under Normal and High Glucose Conditions

Abstract

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.

Graphene Biodevices for Early Disease Diagnosis Based on Biomarker Detection

“…Nitric oxide (NO) is a free radical for crucial transcellular messengers. It plays a significant role in multiple physiological and pathological processes, such as blood pressure regulation, mitochondrial respiration, immune reactions, and vasodilation. − …”

Differentiated PDGFRα-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase

“…Interestingly, low levels of NO were detected in the control conditions suggesting that PDGFRα-positive cells release NO in a spontaneous manner. This is not a new concept as previous studies have demonstrated NO production from human dermal fibroblasts under normal physiological conditions [ 48 ]. In agreement with previous findings that demonstrated that NO is a survival factor for ICC, the data presented in this study have shown that the survival rate of PDGFRα-positive cells was enhanced in response to NO [ 22 ].…”

A Case Report of Exacerbation of Leg Ulcers Associated with Acute High-dose Acetylsalicylic Acid in a Patient with Klinefelter Syndrome

“…The proper concentration of gaseous nitric oxide at the location of wounded tissue seems to be essential for ulcer healing [17]. Nitric oxide is a potent cellular signaling mediator and has been reported to exert both beneficial and pathological effects, depending on the delivered dose and activation/inhibition of the nitric oxide-dependent cellular signaling pathways.…”

“…Nitric oxide is a potent cellular signaling mediator and has been reported to exert both beneficial and pathological effects, depending on the delivered dose and activation/inhibition of the nitric oxide-dependent cellular signaling pathways. For instance, insufficient production of nitric oxide is a major contributor to the dysregulation of wound healing in diabetic foot ulcer [17]. Recent reports suggest it is plausible to assume that there is a dose-dependent effect of acetylsalicylic acid on testosterone level which per se is a regulator of nitric oxide production, signaling, and turnover [18,19].…”