Investigative Study on Nitric Oxide Production in Human Dermal Fibroblast Cells under Normal and High Glucose Conditions (original) (raw)
Related papers
Nitric oxide enhances experimental wound healing in diabetes
British Journal of Surgery, 2002
Background Diabetes is characterized by a nitric oxide deficiency at the wound site. This study investigated whether exogenous nitric oxide supplementation with the nitric oxide donor molsidomine (N-ethoxycarbomyl-3-morpholinyl-sidnonimine) could reverse the impaired healing in diabetes. Methods Wound healing was studied by creating a dorsal skin incision with subcutaneous polyvinyl alcohol sponge implantation in diabetic and non-diabetic rats. Half of each group was treated with molsidomine. Collagen metabolism was assessed by wound breaking strength, hydroxyproline (OHP) content, RNA expression for collagen type I and III, and matrix metalloproteinase (MMP) 2 activity in wound sponges. Wound fluid, plasma and urinary nitric oxide metabolite levels, and the number of inflammatory cells were assessed. Results OHP content and wound breaking strength were significantly increased by molsidomine. MMP-2 activity in wound fluid was decreased in diabetes and upregulated by nitric oxide don...
Therapeutic Role of Nitric Oxide in Diabetic Wound Healing: A Systematic Review
Journal of Pharmaceutical Research International, 2021
Post injury, healing of wound is essential for recovery of uprightness of the body, which is one of the complex, continuous and unanticipated chains of events in case of diabetic patients. Nitric oxide represents a potential wound therapeutic agent due to its ability to regulate inflammation and eradicate bacterial infections. Impaired wound healing is a prominent diabetic complication which may lead to amputations also. In addition to modern medicines we can use nitric oxide therapy prominently for diabetic wound healing. Prominent and proven role of nitric oxide as well as conventional materials (like metformin and hydrogen sulphide, whey proteins, acidified nitrile etc), therapies (like low level laser therapy, hyperbaric oxygen therapy etc) and techniques (like in vivo implants with biosensors) can be taken into consideration. Many plant extracts showed promising results for wound healing activity by increasing nitric oxide levels. Use of modern technologies such as implant with...
Diabetologia, 1999
In diabetic foot ulcers (DFUs) the normal sequence of events in the healing process is disrupted. The condition is a major clinical concern, both in terms of cost ($500 million per year [1]), time and debility for the patient. It is the commonest cause of admission to hospital amongst diabetic patients [2] resulting in 50±70 % of all non-traumatic amputations [3] with a cost of at least £ 7000 per case in the United Kingdom [4]. Furthermore, with an increasingly aged population there is likely to be a rise in the incidence of these ulcers. It has therefore been the goal of clinicians for years to stimulate the normal reparative process in this debilitating disorder. The severe tissue destruction associated with DFUs suggests a number of candidate factors in their aetiology, in particular the metabolism of l-arginine is vital in the repair process. The metabolism of larginine by the enzyme nitric oxide synthase (NOS) Diabetologia (1999) 42: 748±757
Iranian Journal of Veterinary Medicine, 2010
Exogenous nitric oxide donors such as DETA NONOate, spontaneously release nitric oxide. This study aimed to investigate the effect of DETA NONOate as a nitric oxide releasing drug on the rate of collagen synthesis during the impaired wound healing in a rat model of diabetes. Twelve male Sprague–Dawley rats were transferred into separate metabolic cages. Nine days before wounding, the rats were injected intraperitoneally with streptozotocin (STZ; 55 mg/kg body weight in citrate buffer 0.1 mol/L, pH 4.5) to induce diabetes. The dorsal surface of each rat was properly shaved and a full thickness dermal wound was made. The test group (n=6) was treated with 100 ?M DETA NONOate in phosphate buffer while the control wounds (n=6) received sterile saline (PBS) only on the same day as wounding and every three days for one week. After the skin incision, polyvinyl alcohol (PVA) sponges were implanted subcutaneously on the dorsal of each animal under sterile conditions for the collection of woun...
Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society, 2018
The aim of this multi-center, prospective, observer-blinded, parallel group, randomized controlled trial was to assess the safety and efficacy of EDX110, a nitric oxide generating medical device, in the treatment of diabetic foot ulcers in a patient group reflecting "real world" clinical practice compared against optimal standard care. Participants were recruited from ten hospital sites in multidisciplinary foot ulcer clinics. The ulcers were full thickness, with an area of 25-2500mmand either a palpable pedal pulse or ankle brachial pressure index >0.5. Infected ulcers were included. Treatment lasted 12 weeks, or until healed, with a 12-week follow-up period. Both arms were given optimal debridement, offloading and antimicrobial treatment, the only difference being the fixed used of EDX110 as the wound dressing in the EDX110 group. 135 participants were recruited with 148 ulcers (EDX110 - 75; Control - 73), 30% of which were clinically infected at baseline. EDX110 achi...
The effects of topical treatment with acidified nitrite on wound healing in normal and diabetic mice
Nitric Oxide, 2006
The eVects of the application of a nitric oxide generating acidiWed nitrite cream comprising sodium nitrite and citric acid, on the healing of incisional wounds in mice, has been investigated. The eVects of acidiWed nitrite on wound healing were critically dependent on the time of application after wounding. Application of acidiWed nitrite starting on the day of wounding and on consecutive days thereafter signiWcantly inhibited both half time to closure and extent of wound closure. Conversely, application starting on days 1-4 after wounding and on consecutive days thereafter signiWcantly augmented the rate and extent of wound healing. Optimal eVects on improving wound healing were observed with cream concentrations of 3.0% (w/v) sodium nitrite and 4.5% (w/v) citric acid. Starting application on day 5 after wounding had no eVect on the rate or extent of wound healing. In diabetic Lepr db/db mice, starting treatment at day 2 after wounding, acidiWed nitrite at 3.0% (w/v) sodium nitrite and 4.5% (w/v) citric acid signiWcantly increased the rate and extent of wound healing. This suggests that acidiWed nitrite is eVective in improving wound healing against a diabetic background. The present data shows that acidiWed nitrite cream, a clinically eVective means of topically delivering nitric oxide, augments the wound healing process and may be of clinical beneWt.
Nitric oxide synthesis inhibition alters rat cutaneous wound healing
Journal of Cutaneous Pathology, 2006
Background: Nitric oxide (NO) is an important molecule that participates in wound repair, but its effects on cutaneous wound healing are not well understood. The aim of this study was to investigate the effects of NO synthesis blockade on rat cutaneous wound healing by the administration of NG‐nitro‐l‐arginine methyl ester (l‐NAME), a non‐selective inhibitor of NO synthases.Methods: NO synthesis was inhibited by administration of l‐NAME (20 mg/kg/day) in drinking water. An excisional wound was done, and the animals were killed 7, 14, and 21 days later. Wound contraction and blood pressure were evaluated. The lesion and adjacent skin were formalin fixed and paraffin embedded. Mast cells were quantified, and vessels were evaluated using stereological methods.Results: l‐NAME‐treated animals presented delayed wound contraction, alterations in collagen organization, and neoepidermis thickness. The inhibition of NO synthesis increased mast cell migration 7 days after wounding, but decr...
Nitric Oxide Metabolism in Wounds
Journal of Surgical Research, 1997
Arginine can be metabolized in wounds to nitric oxide and citrulline by nitric oxide synthase or to urea and ornithine by arginase. We investigated the expression of these arginine metabolic pathways over a 3-week period. Groups of 8-10 male Balb/C mice underwent a dorsal skin incision and subcutaneous polyvinyl alcohol sponge implantation. The animals were sacrificed at various times, and sponges were harvested to obtain wound fluid and wound cells. Cells or whole sponges were incubated with L-[2,3-(3)H]arginine, with or without N(G)-L-monomethyl-arginine (NMMA, a competitive inhibitor of nitric oxide synthase). Nitrite and nitrate (both stable end products of nitric oxide metabolism) and amino acids were measured in wound fluid and wound cell culture supernatants. Increasing concentrations of nitrite and nitrate were noted in wound fluid and in whole sponge cultures until the second week postwounding, indicating sustained wound nitric oxide synthesis. In wound fluid arginine levels were undetectable at all times, suggesting sustained utilization. Wound fluid citrulline levels showed an early peak and then a gradual decrease, suggesting that recycling for continued nitric oxide production may occur. Wound fluid ornithine levels increased until Day 10 and remained elevated, indicative of continued arginase activity. In vitro production of nitrite/nitrate and citrulline by cells and whole sponges was inhibitable by NMMA. Inducible nitric oxide synthase expression was confirmed by immunoblotting, while immunohistochemistry demonstrated that macrophages are a major source of wound nitric oxide. The data show that nitric oxide synthesis occurs for prolonged periods after injury and macrophages appear to be a major cellular source.
Wound Repair and Regeneration, 2002
Healing of chronic wounds such as diabetic foot ulcers is a significant clinical problem. Methods of accelerating healing in these difficult lower extremity sites include use of growth factor-loaded gels, hyperbaric oxygen, grafts, and artificial skin replacements. Nitric oxide (NO) has been proposed as a possible active agent for enhancing wound healing. This study examines the in vitro and in vivo responses to a novel hydrogel that produces therapeutic levels of NO. A hydrogel wound dressing was fabricated using ultraviolet light-initiated polymerization from poly(vinyl alcohol) with a NO donor covalently coupled to the polymer backbone. NO release from the NO-modified hydrogel was shown to occur over a time period of up to 48 hours, and there was no associated decrease in fibroblast growth or viability in vitro associated with NO hydrogels. Fibroblasts in culture with NO hydrogels had an increased production of extracellular matrix compared with cells cultured without the NO hydrogels. Preliminary animal studies in a diabetic mouse, impaired wound healing model were conducted comparing low (0.5 mM) and high (5 mM) doses of NO. Time to complete closure was similar in control wounds and NO-treated wounds; however, at 8 days control wounds were significantly smaller than NO-treated wounds. By days 10 to 13 this delay was no longer apparent. Granulation tissue thickness within the wounds at days 8 and 15 and scar tissue thickness after wound closure were increased in animals exposed to higher dose NO hydrogels. The results of this study suggest that exogenous NO released from a hydrogel wound dressing has potential to modulate wound healing. (WOUND REP REG 2002;10:286-294) Chronic wounds such as pressure ulcers, diabetic foot ulcers, and venous ulcers make up approximately 70% of all skin wounds. 1 Chronic wounds like these incur a huge cost From the Departments of Chemical Engineering
FEBS Letters, 2000
Glucose-6-phosphate dehydrogenase (G6PD) plays an important role in cellular redox homeostasis, which is crucial for cell survival. In the present study, we found that G6PD status determines the response of cells exposed to nitric oxide (NO) donor. Treatment with NO donor, sodium nitroprusside (SNP), caused apoptosis in G6PD-deficient human foreskin fibroblasts (HFF1), whereas it was growth stimulatory in the normal counterpart (HFF3). Such effects were abolished by NO scavengers like hemoglobin. Ectopic expression of G6PD in HFF1 cells switched the cellular response to NO from apoptosis to growth stimulation. Experiments with 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one and 8-bromo-cGMP showed that the effects of NO on HFF1 and HFF3 cells were independent of cGMP signalling pathway. Intriguingly, trolox prevented the SNP-induced apoptosis in HFF1 cells. These data demonstrate that G6PD plays a critical role in regulation of cell growth and survival. ß