Mitochondrial Redox Balance of Fibroblasts Exposed to Ti-6Al-4V Microplates Subjected to Different Types of Anodizing.

Despite the high biocompatibility of titanium and its alloys, the need to remove titanium implants is increasingly being debated due to the potential for adverse effects associated with long-term retention. Therefore, new solutions are being sought to enhance the biocompatibility of titanium implants. One of them is to increase the thickness of the passive layer of the implant made of titanium dioxide. We were the first to evaluate the effect of hard-anodized (type II) Ti-6Al-4V alloy discs on the cytotoxicity, mitochondrial function, and redox balance of fibroblasts mitochondria compared to standard-anodized (type III) and non-anodized discs. The study used fibroblasts obtained from human gingival tissue. The test discs were applied to the bottom of 12-well plates. Cells were cultured for 24 h and 7, 14, and 21 days and mitochondria were isolated. We demonstrated the occurrence of oxidative stress in the mitochondria of fibroblasts of all tested groups, regardless of the presence and type of anodization. Type II anodization prevented changes in complex II activity (vs. control). The lowest degree of citrate synthase inhibition occurred in mitochondria exposed to titanium discs with type II anodization. In the last phase of culture, the presence of type II anodization reduced the degree of cytochrome c oxidase inhibition compared to the other tests groups and the control group, and prevented apoptosis. Throughout the experiment, the release of titanium, aluminium, and vanadium ions from titanium discs with a hard-anodized passive layer was higher than from the other titanium discs, but decreased with time. The obtained results proved the existence of dysfunction and redox imbalance in the mitochondria of fibroblasts exposed to hard-anodized titanium discs, suggesting the need to search for new materials perhaps biodegradable in tissues of the human body.

Can smoking alter salivary homeostasis? A systematic review on the effects of traditional and electronic cigarettes on qualitative and quantitative saliva parameters.

The available literature indicates that smoking causes quantitative and qualitative changes in saliva. However, there is a lack of studies summarizing the knowledge in this area, and there are no clear guidelines on the use of salivary biomarkers for assessing exposure to cigarette smoke (CS). The present work aimed to provide a systematic review of the literature regarding the influence of smoking traditional and electronic cigarettes, as well as heat-not-burn products, on salivary homeostasis. An electronic search of the literature from 1982 to 2023 was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Based on the inclusion criteria, 65 studies were used for the final review. Smoking traditional as well as electronic cigarettes negatively affects salivary biomarkers, including the salivary flow rate, pH, antibody titer, electrolyte concentration, microflora composition, redox balance, and inflammation, in terms of both quantity and quality. However, to date, only single salivary biomarkers have been compared in traditional and electronic cigarette smokers. It can be concluded that the salivary production rate, pH, microbiome, and cytokines can be used to assess exposure to CS smoke. There is a lack of convincing evidence to compare the toxic influence of traditional and electronic cigarettes on salivary homeostasis. Future experiments should include long-term randomized clinical trials on larger populations of smokers.

Significance of nitrosative stress and glycoxidation products in the diagnosis of COVID-19.

Nitrosative stress promotes protein glycoxidation, and both processes can occur during an infection with the SARS-CoV-2 virus. Therefore, the aim of this study was to assess selected nitrosative stress parameters and protein glycoxidation products in COVID-19 patients and convalescents relative to healthy subjects, including in reference to the severity of COVID-19 symptoms. The diagnostic utility of nitrosative stress and protein glycoxidation biomarkers was also evaluated in COVID-19 patients. The study involved 218 patients with COVID-19, 69 convalescents, and 48 healthy subjects. Nitrosative stress parameters (NO, S-nitrosothiols, nitrotyrosine) and protein glycoxidation products (tryptophan, kynurenine, N-formylkynurenine, dityrosine, AGEs) were measured in the blood plasma or serum with the use of colorimetric/fluorometric methods. The levels of NO (p = 0.0480), S-nitrosothiols (p = 0.0004), nitrotyrosine (p = 0.0175), kynurenine (p < 0.0001), N-formylkynurenine (p < 0.0001), dityrosine (p < 0.0001), and AGEs (p < 0.0001) were significantly higher, whereas tryptophan fluorescence was significantly (p < 0.0001) lower in COVID-19 patients than in the control group. Significant differences in the analyzed parameters were observed in different stages of COVID-19. In turn, the concentrations of kynurenine (p < 0.0001), N-formylkynurenine (p < 0.0001), dityrosine (p < 0.0001), and AGEs (p < 0.0001) were significantly higher, whereas tryptophan levels were significantly (p < 0.0001) lower in convalescents than in healthy controls. The ROC analysis revealed that protein glycoxidation products can be useful for diagnosing infections with the SARS-CoV-2 virus because they differentiate COVID-19 patients (KN: sensitivity-91.20%, specificity-92.00%; NFK: sensitivity-92.37%, specificity-92.00%; AGEs: sensitivity-99,02%, specificity-100%) and convalescents (KN: sensitivity-82.22%, specificity-84.00%; NFK: sensitivity-82,86%, specificity-86,00%; DT: sensitivity-100%, specificity-100%; AGE: sensitivity-100%, specificity-100%) from healthy subjects with high sensitivity and specificity. Nitrosative stress and protein glycoxidation are intensified both during and after an infection with the SARS-CoV-2 virus. The levels of redox biomarkers fluctuate in different stages of the disease. Circulating biomarkers of nitrosative stress/protein glycoxidation have potential diagnostic utility in both COVID-19 patients and convalescents.

Redox Biomarkers - An Effective Tool for Diagnosing COVID-19 Patients and Convalescents.

Aim: COVID-19 triggers the overproduction of reactive oxygen species (ROS) which, in combination with a weakened antioxidant barrier, can lead to protein oxidation and lipid peroxidation. The aim of this study was to evaluate enzymatic and non-enzymatic antioxidants, the overall redox potential, and protein and lipid peroxidation products in COVID-19 patients, convalescents, and healthy subjects, and to the determine the diagnostic applicability of these parameters in COVID-19 patients. Materials and Methods: The study involved 218 patients with COVID-19, 69 convalescents, and 48 healthy subjects who were selected for the research based on age and sex. The study was conducted between 20 February 2021 and 20 November 2021 in Bialystok, Poland. The antioxidant barrier, redox status, and oxidative damage products were assessed in serum/plasma samples with the use of colorimetric and spectrophotometric assays. Results: Glutathione reductase (GR) activity was higher, whereas total antioxidant capacity (TAC) was lower in COVID-19 patients than in convalescents (p<0.0001) and the control group (p<0.0001). The concentrations of advanced glycation end products (AGEs), advanced oxidation protein products (AOPP), 4-hydroxynonenal (4-HNE), and malondialdehyde (MDA) were higher in COVID-19 patients (p<0.0001) and convalescents (p<0.0001) than in the control group. AGEs were the most effective diagnostic biomarker for differentiating COVID-19 patients from the control group (AUC=0.9971) and convalescents from the control group (AUC=1.000). Conclusion: An infection with the SARS-CoV-2 disrupts the redox balance and increases protein oxidation and lipid peroxidation. AGEs fulfill the criteria for a potential diagnostic biomarker in COVID-19 patients and convalescents.

Could circulating biomarkers of nitrosative stress and protein glycoxidation be useful in patients with gastric cancer?

Background: Nitrosative stress leads to protein glycoxidation, but both processes may be strongly related to the cancer development. Therefore, the aim of this study was to assess the nitrosative stress and protein glycoxidation products in patients with gastric cancer in comparison with healthy controls. We are also the first to evaluate the diagnostic utility of nitrosative stress and protein glycoxidation markers in gastric cancer patients in respect to histopathological classifications (TNM, Lauren's and Goseki's classification) and histopathological parameters such as histological type, histological differentiation grade, presence of vascular or neural invasion, desmoplasia and Helicobacter pylori infection. Methods: The study included 50 patients with gastric cancer and 50 healthy controls matched for sex and age. Nitrosative stress parameters and protein glycoxidation products were measured colorimetrically/fluorometrically in plasma or serum samples. Student's t-test or Mann-Whitney U-test were used for statistical analysis. Results: NO, S-nitrosothiols, nitrotyrosine, kynurenine, N-formylkynurenine, dityrosine, AGE and Amadori products were significantly increased whereas tryptophan fluorescence was decreased in patients with gastric cancer compared to the healthy control. Nitrosative stress and glycoxidation products may be useful in diagnosis of gastric cancer because they differentiate patients with gastric cancer from healthy individuals with high sensitivity and specificity. Some of the determined parameters are characterised by high AUC value in differentiation of GC patients according to the histopathological parameters. Conclusions: Gastric cancer is associated with enhanced circulating nitrosative stress and protein glycation. Although further research on a tissue model is needed, plasma/serum biomarkers may be dependent on tumour size, histological type, tumour invasion depth, presence of lymph node and distant metastasis, vascular and neural invasion and Helicobacter pylori infection. Thus, circulating biomarkers of nitrosative stress/protein glycoxidation may have potential diagnostic significance in gastric cancer patients.

Diagnostic significance and utility of circulating redox biomarkers in patients with gastric cancer - preliminary study.

BACKGROUND: This study aimed to evaluate the redox status, antioxidant barrier, and oxidative damage to proteins, lipids, and DNA in patients with gastric cancer (GC). We are also the first to assess the diagnostic utility of redox parameters in patients with GC with respect to histopathological parameters. METHODS: Fifty patients with gastric cancer and 50 healthy controls matched for sex and age were included in the study. The antioxidant barrier, redox status, and oxidative damage products were measured in serum/plasma samples using colorimetric or spectrophotometric methods. RESULTS: The activity of superoxide dismutase - SOD (p < 0.05) was significantly higher, whereas the activities of catalase - CAT (p < 0.0001), glutathione peroxidase - GPx (p < 0.0001), glutathione reductase - GR (p < 0.0001), and reduced glutathione - GSH (p < 0.05) were considerably lower in GC patients than in the control group. The levels of total oxidant status - TOS (p < 0.0001), oxidative stress index - OSI (p < 0.0001), advanced oxidation protein products - AOPP (p < 0.0001), ischaemia modified albumin - IMA (p < 0.01), lipid hydroperoxides - LOOH (p < 0.0001), 8-IsoProstane - 8-Iso-P (p < 0.0001), and DNA/RNA (p < 0.0001) were significantly higher, and the levels of total antioxidant capacity - TAC (p < 0.0001) and total thiols (p < 0.0001) were considerably lower in patients compared to the healthy controls. Some redox parameters are characterized by high AUC values in patients with differentiated GC according to histopathological parameters. CONCLUSIONS: Gastric cancer is strongly linked to a systemic redox imbalance and increased oxidative damage to proteins, lipids, and DNA. Redox biomarkers are potential diagnostic indicators of gastric cancer advancement.

Gastric cancer is associated with redox imbalance and increased oxidative damage to proteins, lipids and DNA.Histopathological parameters of the tumour, such as size, histological type, histological differentiation grade, tumour invasion depth, presence of lymph node and distant metastasis, Lauren and Goseki classification, and presence of vascular and neural infiltration, are associated with the level of antioxidants and oxidative damage products of proteins, lipids, and DNA.Determination of redox parameters may be useful in the assessment of the tumour progression.

Sources of free radicals and oxidative stress in the oral cavity.

OBJECTIVE: An oral cavity is a place especially susceptible to oxidative damage. It is subjected to many environmental pro-oxidative factors or factors that have the ability to generate reactive oxygen species (ROS). The aim of this article is to present the main sources of ROS and oxidative stress in the oral environment. DESIGN: A literature search was performed using the PubMed and Google Scholar databases. RESULTS: One of the most important ROS sources in the oral cavity is periodontal inflammation. Other sources of ROS include: xenobiotics (ethanol, cigarette smoke, drugs), food (high-fat diet, high-protein diet, acrolein), dental treatment (ozone, ultrasound, non-thermal plasma, laser light, ultraviolet light), and dental materials (fluorides, dental composites, fixed orthodontic appliances, and titanium fixations). It has been shown that excessive production of ROS in the oral cavity may cause oxidative stress and oxidative damage to cellular DNA, lipids, and proteins, thus predisposing to many oral and systemic diseases. CONCLUSIONS: Recognition of the exogenous sources of ROS and limitation of exposure to the ROS generating factors can be one of the prophylactic measures preventing oral and systemic diseases. It is suggested that antioxidant supplementation may be helpful in people exposed to excessive production of ROS in the oral cavity system.

Effect of N-Acetylcysteine on Antioxidant Defense, Oxidative Modification, and Salivary Gland Function in a Rat Model of Insulin Resistance.

Oxidative stress plays a crucial role in the salivary gland dysfunction in insulin resistance (IR). It is not surprising that new substances are constantly being sought that will protect against the harmful effects of IR in the oral cavity environment. The purpose of this study was to evaluate the effect of N-acetylcysteine (NAC) on oxidative stress and secretory function of salivary glands in a rat model of insulin resistance. Rats were divided into 4 groups: C-normal diet, C + NAC-normal diet + NAC, HFD-high-fat diet, and HFD + NAC. We have demonstrated that NAC elevated enzymatic (superoxide dismutase, catalase, and peroxidase) and nonenzymatic antioxidants (reduced glutathione (GSH) and total antioxidant capacity (TAS)) in the parotid glands of HFD + NAC rats, while in the submandibular glands increased only GSH and TAS levels. NAC protects against oxidative damage only in the parotid glands and increased stimulated salivary secretion; however, it does not increase the protein secretion in the both salivary glands. Summarizing, NAC supplementation prevents the decrease of stimulated saliva secretion, seen in the HFD rats affected. NAC improves the antioxidative capacity of the both glands and protects against oxidative damage to the parotid glands of IR rats.

Chronic high-protein diet induces oxidative stress and alters the salivary gland function in rats.

OBJECTIVE: Chronic high protein intake leads to an increase in reactive oxygen species (ROS) generation. However, there is no data on the impact of high-protein diet on the antioxidant barrier, oxidative stress and secretory function in the salivary glands of healthy individuals. DESIGN: 16 male Wistar rats were randomly divided into 2 groups (n=8): normal protein (C) and high-protein diet (HP) for 8 weeks. Salivary antioxidants: peroxidase (Px), catalase (CAT), superoxide dismutase 1 (SOD 1), uric acid (UA), total antioxidant status (TAS), total oxidant status (TOS) and the oxidative stress index (OSI), as well as protein carbonyls (PC), 4-hydroxynonenal protein adduct (4-HNE protein adduct), 8-isoprostanes (8-isoP), 8-hydroxy-2'-deoxyguanosine (8-OHdG) and protein content were determined in the salivary glands and plasma. Salivary unstimulated and stimulated flow rates were examined. RESULTS: Parotid Px, TAS, UA, TOS, OSI, PC were significantly higher, the total protein content was statistically lower in the HP group as compared to the control. Submandibular UA, TOS, OSI, 8-isoP, 4-HNE-protein adduct, 8-OHdG were statistically elevated, SOD 1 and Px were significantly lower in the HP group as compared to the control rats. The unstimulated salivary flow rate was significantly depressed in the HP group as compared to the controls. CONCLUSIONS: Higher antioxidant capacity in the parotid glands of HP rats vs. control rats seems to be a response to a higher ROS formation. In the submandibular glands severe oxidative modification of almost all cellular components was observed. Administration of HP resulted in the weakening of the salivary gland function.

Oxidative Modification in the Salivary Glands of High Fat-Diet Induced Insulin Resistant Rats.

Still little is known about the role of oxidative stress (OS) in the pathogenesis of the salivary gland dysfunction in the course of insulin resistance (IR). To induce IR rats was fed with a high fat diet (HFD) during 8 weeks. Stimulated and non-stimulated salivary flow rate, total protein, as well as oxidative damage markers: 4-HNE protein adduct, 8-isoprostanes (8-isoP), 8-hydroxy-D-guanosine (8-OHdG), advanced oxidation protein product (AOPP), and protein carbonyls (PC) were determined in the plasma and submandibular and parotid glands of IR and control rats. We have shown a significant decrease (45%) of the stimulated salivary flow rate, and in the total protein concentration in the parotid (35%) and submandibular (10%) glands of HFD IR as compared to the control rats. The level of 4-HNE protein adduct (15%) and 8-isoP (20%) in the submandibular glands of IR rats as well as total level of 4-HNE protein adduct (39%), 8-isoP (27%), AOPP (25%), PC (32%), and 8-OHdG (18%) in the parotid glands of IR rats were significantly higher as compared to the control group. We showed no correlation between the assessed OS parameters in the plasma and salivary glands. However, the redox balance in both glands shifted toward the oxidative status, parotid glands of IR rats are exposed to greater intensity OS. Stimulated secretory ability and mechanisms involved in the synthesis/secretion of proteins in the salivary glands are depressed in the course of IR. Oxidative damage in the salivary glands arises independently from the general OS in the course of insulin resistance induced by a high fat diet.

The Redox Balance in Erythrocytes, Plasma, and Periosteum of Patients with Titanium Fixation of the Jaw.

Titanium miniplates and screws are commonly used for fixation of jaw fractured or osteotomies. Despite the opinion of their biocompatibility, in clinical practice symptoms of chronic inflammation around the fixation develop in some patients, even many years after the application of miniplates and screws. The cause of these complications is still an unanswered question. Taking into account that oxidative stress is one of the toxic action of titanium, we have evaluated the antioxidant barrier as well as oxidative stress in the erythrocytes, plasma and periosteum covering the titanium fixation of the jaw. The study group was composed of 32 patients aged 20-30 with inserted miniplates and screws. The antioxidant defense: catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase-1 (SOD1), uric acid (UA), total antioxidant capacity (TAC), as well as oxidative damage products: advanced oxidation protein products (AOPP), advanced glycation end products (AGE), dityrosine, kynurenine, N-formylkynurenine, tryptophan, malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), total oxidant status (TOS), and oxidative status index (OSI) were evaluated. SOD1 activity (↓37%), and tryptophan levels (↓34%) showed a significant decrease while AOPP (↑25%), TOS (↑80%) and OSI (↑101%) were significantly elevated in maxillary periosteum of patients who underwent bimaxillary osteotomies as compared to the control group. SOD-1 (↓55%), TAC (↓58.6%), AGE (↓60%) and N-formylkynurenine (↓34%) was statistically reduced while AOPP (↑38%), MDA (↑29%), 4-HNE (↑114%), TOS (↑99%), and OSI (↑381%) were significantly higher in the mandibular periosteum covering miniplates/screw compared with the control tissues. There were no correlations between antioxidants and oxidative stress markers in the periosteum of all patients and the blood. As exposure to the Ti6Al4V titanium alloy leads to disturbances of redox balance in the periosteum surrounding titanium implants of the maxilla and the mandible so antioxidant supplementation should be recommended to the patients undergoing treatment of dentofacial deformities with the use of titanium implants. The results we obtained may also indicate a need to improve the quality of titanium jaw fixations through increase of TiO2 passivation layer thickness or to develop new, the most highly biodegradable materials for their production.