High glucose-induced oxidative stress impairs proliferation and migration of human gingival fibroblasts

Abstract

Delayed gingival wound healing is widely observed in periodontal patients with diabetes. However, the molecular mechanisms of the impaired function of gingival fibroblasts in diabetes remain unclear. The purpose of this study was to investigate changes in the properties of human gingival fibroblasts (HGFs) under high-glucose conditions. Primary HGFs were isolated from healthy gingiva and cultured with 5.5, 25, 50, and 75 mM glucose for 72 h. In vitro wound healing, 5-ethynyl-2′-deoxyuridine (EdU), and water-soluble tetrazolium salt (WST-8) assays were performed to examine cell migration and proliferation. Lactase dehydrogenase (LDH) levels were measured to determine cytotoxicity. The mRNA expression levels of oxidative stress markers were quantified by real-time PCR. Intracellular reactive oxygen species (ROS) were also measured in live cells. The antioxidant N-acetyl-l-cysteine (NAC, 1 mM) was added to evaluate the involvement of ROS in the glucose effect on HGFs. As a result, the in vitro wound healing assay showed that high glucose levels significantly reduced fibroblast migration and proliferation at 6, 12, 24, 36, and 48 h. The numbers of cells positive for EdU staining were decreased, as was cell viability, at 50 and 75 mM glucose. A significant increase in LDH was proportional to the glucose concentration. The mRNA levels of heme oxygenase-1 and superoxide dismutase-1 and ROS levels were significantly increased in HGFs after 72 h of exposure to 50 mM glucose concentration. The addition of NAC diminished the inhibitory effect of high glucose in the in vitro wound healing assay. The results of the present study show that high glucose impairs the proliferation and migration of HGFs. Fibroblast dysfunction may therefore be caused by high glucose-induced oxidative stress and may explain the delayed gingival wound healing in diabetic patients.

High-glucose Induces Retinal Pigment Epithelium Mitochondrial Pathways of Apoptosis and Inhibits Mitophagy by Regulating ROS/PINK1/Parkin Signal Pathway

“…Previous studies have reported that glucose induces ROS generation [14] and inhibits cell proliferation [21]. We first treated RPE with 15uM, 30uM, 50uM and 70uM glucose for 24h, 48h, 72h and 96h respectively, the cell counting and CCK-8 assay results showed that glucose inhibits RPE proliferation in a dose dependent manner (Figure 1A-D).…”

Regenerative and protective effects of calcium silicate on senescent fibroblasts induced by high glucose

“…Substantial evidence suggests that a high level of ROS impairs cell function and induces premature senescence in cells . The increased ROS production accelerates telomere shortening, induces the DNA damage response (DDR), and causes senescence growth arrest .…”

Role of Sex Hormones And Their Receptors On Gastric Nrf2 And Neuronal Nitric Oxide Synthase Function In An Experimental Hyperglycemia Model

“…Furthermore, recent studies have documented that hyperosmolarity, as occurring in diabetic hyperglycemia, may represent important regulatory elements in uencing cell fate and viability, both in physiological and pathological conditions. These studies further report using glucose concentrations between 24mM -75 mM with incubation times up to 72 hr are su cient to mimic the diabetic oxidative stress response in different cellular types (human gingival broblasts and erythrocytes) [39,43]. Furthermore, exposure to a prolonged severe hyperglycemic (>30mM) load is correlated with increased susceptibility to cellular damage and severe inhibitory effects on nNOS/Nrf2.…”