Neopterin, the Cell-Mediated Immune Response Biomarker, in Inflammatory Periodontal Diseases: A Narrative Review of a More than Fifty Years Old Biomarker.

Neopterin is a biomarker of the activation of cellular immunity. The purpose of this review is to summarise neopterin metabolism, methods of its detection, and its role in inflammation, focusing on periodontal inflammatory diseases. This derivative of guanosine is a non-enzymatic product of 7,8-dihydroneopterin oxidation caused by free radicals which protect activated macrophages from oxidative stress. Various methods, usually based on enzyme-linked immunosorbent essay, high-performance liquid chromatography, or radioimmunoassay were developed for the isolation of neopterin. A wide spectrum of diseases and conditions are known to affect neopterin levels, including cardiovascular, bacterial, viral, and degenerative diseases, as well as malignant tumours. Neopterin levels were found to increase in subjects with periodontitis, especially when the oral fluid and gingival crevicular fluid were evaluated. These findings confirm the role of activated macrophages and cellular immunity in periodontal inflammatory diseases. The gingival crevicular fluid and the oral fluid appear to be the most valuable biologic fluids for the evaluation of neopterin levels in periodontitis. For gingival crevicular fluid, neopterin can be determined as the concentration or the so-called total amount. Nonsurgical periodontal treatment was associated with a decrease in neopterin levels, but an increase was also reported, suggesting the possible role of macrophages in the resolution of the periodontal lesion.

Neopterin Levels in Periodontitis and after Nonsurgical Periodontal Therapy: Evaluation of Gingival Crevicular Fluid, Oral Fluid, Serum and Urinary Samples-A Case-Control Study.

Periodontitis is a chronic inflammatory disease that leads to the destruction of the tooth-supporting tissues with complex immune response. Neopterin (Np), secreted via activated macrophages, is considered a biomarker of cellular immunity. The aim of this study was to evaluate the impact of periodontitis and nonsurgical periodontal therapy. Np gingival crevicular fluid (GCF), oral fluid, serum and urine levels were compared in subjects with periodontitis before periodontal treatment, three months after and in a healthy control. Np GCF concentrations in the study group after treatment were significantly higher than the control group (p = 0.038). The GCF total amount (amount of substance) was significantly higher in the study group before periodontal treatment than in the control group (p = 0.001) and higher than the levels taken after treatment collection (p = 0.024). The oral fluid Np concentrations in the study group after treatment were significantly increased compared to the before treatment concentrations (p = 0.020). The same trend was observed in the urine samples. Significant correlation was found between the serum and oral fluid Np concentrations (p = 0.001, ρ = 0.40). Our results confirm the impact of cellular immunity and macrophages on periodontitis and on the resolution of periodontal inflammation. The presence of neopterin in oral fluid most likely originates in the serum.

Liquid chromatography method with tandem mass spectrometry and fluorescence detection for determination of inflammatory biomarkers in gingival crevicular fluid as a tool for diagnosis of periodontal disease.

The new ultra-high performance liquid chromatography method with tandem mass spectrometry and fluorescence detection allowing fast, selective, and high-throughput analysis of neopterin, kynurenine, tryptophan, and creatinine in gingival crevicular fluid (GCF) has been optimized. Defining the pathophysiology of periodontal disease and identification of potential diagnostic test for active periodontitis remains a significant challenge in the field of oral disease diagnosis. Analysis of GCF provides a non-invasive means of evaluating the role of the host response in periodontal disease. In addition, the analysis of GCF provides an information about current inflammation level of sampled site/tooth. Determination of GCF inflammatory biomarkers such as neopterin, kynurenine, and tryptophan can contribute to diagnosis, evaluation of treatment, and progression of periodontal diseases such as gingivitis and periodontitis. The separation of target analytes was carried out using a column Kinetex™ Polar C18 100 Å, (100 × 3.0 mm) packed with 2.6 µm core-shell particles applying an elution with a gradient formed from 0.2% aqueous formic acid and 90% aqueous acetonitrile. Kynurenine, tryptophan, and creatinine were detected using mass spectrometry with electrospray ionization to improve the sensitivity while neopterin was detected using fluorescence detection. The separation of these four substances was achieved after using a very simple sample preparation technique convenient for small amount of biological sample. Only less than 20 µL sample was needed and the separation was completed in 4 min. MS/MS analysis was performed using multiple reaction monitoring (MRM) under a positive ionization mode. Deuterium labeled internal standard was used for the more precise quantification. The lower limits of quantification (LLOQ) for target analytes were 0.50 × 10-3 µmol/L for neopterin, 0.10 µmol/L for kynurenine, and 0.20 µmol/L for tryptophan and creatinine. The within-run and between-run accuracy were in a range of 96.67-114.77% for all quality controls and LLOQ of all analytes. Matrix effect, extraction recovery, and stability testing have all been investigated. The method was tested with real-life samples using GCF collected from patients suffering from periodontitis and from healthy controls. Neopterin levels in patients were significantly higher (P = 0.020) than in healthy subjects and indicate good potential of this method for using in evaluation of periodontal pathogenesis and healing outcomes following a treatment.

Oral Mucositis Association with Periodontal Status: A Retrospective Analysis of 496 Patients Undergoing Hematopoietic Stem Cell Transplantation.

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) can induce serious oral complications, including oral mucositis (OM). The presence of periodontal inflammation before HSCT is believed to be associated with OM. The aim of our study was to determine the prevalence and severity of OM in patients undergoing HSCT and its relation to periodontal status. PATIENTS AND METHODS: This is a retrospective study of patients who underwent HSCT and a detailed dental examination between 2007 and 2015. The dental and periodontal status of all patients was evaluated by clinical and radiographic examination prior to HSCT. Oral health was assessed with the gingival index, the the community periodontal index, presence of plaque-related gingivitis, and marginal periodontitis. During the HSCT period, patients were examined daily for the presence of OM, which was graded according to World Health Organization (WHO) classification if present. The patients were assigned to the groups according to type of transplantation: autologous HSCT, myeloablative allogeneic HSCT, and non-myeloablative allogeneic HSCT. RESULTS: A total of 496 patients were included in the study. OM was present in 314 of 496 patients (63.3%): 184/251 (73.3%) in the autologous group, 100/151 (66.2%) in the myeloablative allogeneic group, and 30/94 (31.9%) in the nonmyeloablative allogeneic group. Significantly more patients suffered from OM in the autologous and myeloablative groups versus the nonmyeloablative conditioning group (p < 0.001). The presence of periodontal inflammation did not significantly differ among the groups. There was only a borderline trend for the higher prevalence of OM in the non-myeloablative allogeneic nonmyeloablative group when periodontal inflammation was present (0.073939). CONCLUSIONS: Oral mucositis prevalence and severity after stem cell transplantation is not widely affected by the oral hygiene and periodontal disease presence before HSCT. We confirmed the wide-known connection of the conditioning regimen intensity to the prevalence of OM.

Analysis of Coexistence of Oral and Cutaneous Lesions in 253 Patients with Lichen Planus - Single-center Retrospective Analysis.

Lichen planus (LP) is a chronic inflammatory disorder that especially affects the skin, mucous membranes, or both. The aim of the study was to determine the clinical characteristics of patients with LP in the oral cavity and concomitant cutaneous lesions and compare their outcomes with those without cutaneous lesions. 253 records of patients with confirmed diagnosis of oral lichen planus (OLP) were retrospectively analyzed. The following clinical data were obtained from the medical charts: sex, age, clinical presentations of OLP, distributions of the lesions, presence of symptoms, extra oral manifestations of lichen planus, presence of systemic diseases, and treatment provided. The group of patients with cutaneous manifestations was compared for possible clinical differences to those without. Cutaneous lesions were present in 18.2% (46/253) of patients. Significantly more patients with cutaneous lesions had other extra oral manifestations (26.1% versus 1.0%, P<0.00001). Lips were significantly more affected in patients with cutaneous presentation (41.3% versus 16.9%, P=0.00006). Ulcerative OLP was more frequent in patients with cutaneous lesions (23.9% versus 10.6%, P=0.0266). Patients with cutaneous manifestations needed significantly more systemic treatment with systemic steroids (10.9% versus 3.4%, P=0.0466). Patients with cutaneous lesions were more symptomatic and tended to require more treatment than patients with OLP who only had oral involvemen.

The Characteristics of Patients with Oral Lichen Planus and Malignant Transformation-A Retrospective Study of 271 Patients.

INTRODUCTION: Oral lichen planus (OLP) is a chronic inflammatory disease with an unknown etiology rating among oral potentially malignant disorder. The aim of the study was to determine the epidemiological and clinical characteristics of the patients with OLP and rate of malignant transformation. PATIENTS AND METHODS: Data were obtained from the medical records of 271 patients referred to the Oral Medicine Unit at the University Hospital in Hradec Králové diagnosed with oral lichen planus in the period of 2003-2020. The records were retrospectively analyzed. The following clinical data were retrieved from the medical charts: gender, age, systemic diseases, alcohol and tobacco consumption, localization/clinical appearance of lesions, distribution of the lesions, presence of the symptoms, treatment provided and malignant transformation. RESULTS: A total of 271 charts of patients with confirmed diagnosis of OLP were retrospectively analyzed, of whom, 66.4% (180/271) were women and 33.6% (91/271) were men. The mean age of the patients was 56.0 (18.2-85.0) years. The median follow-up of all patients was 15.2 months. Overall, 2 patients (2/271, 0.74%) meeting the above-mentioned criteria for malignant transformation were identified during the follow-up period. Both patients suffered from erosive type OLP and developed squamous cell carcinoma of the tongue. CONCLUSIONS: This retrospective study is in concordance with other studies showing the similar profile and clinical features of the patients with OLP. Malignant transformation rate was 0.74%.

Chromatographic method for the determination of inflammatory biomarkers and uric acid in human saliva.

Determination of concentration of biomarkers of the activation of immune system, uric acid, and creatinine in the saliva can be useful tool for the diagnosis and monitoring of early manifestations of diseases such as malignant, inflammatory, and periodontal disorders. We have developed and validated a high-performance liquid chromatographic method coupled with fluorescence and diode array detection for the separation and quantification of neopterin, tryptophan, creatinine, uric acid, and kynurenine in the human saliva. A separation of these analytes was achieved within 9 min by using second-generation monolithic stationary phase and elution with phosphate buffer. The present method involves very simple sample preparation requiring small amount of sample matrix. The internal standard 3-nitro-l-tyrosine was used for a more precise quantification. The sensitivity of the present method was demonstrated with lower limits of quantification of 0.6 × 10-3 µmol/L for neopterin, 0.725 µmol/L for tryptophan, 0.12 µmol/L for creatinine, 0.18 µmol/L for uric acid, and 0.135 µmol/L for kynurenine. The method was validated with 67 real-life saliva samples collected from patients suffering from breast, ovarian, colorectal, and renal cancer, and 19 saliva samples from patients with periodontal diseases and allowed monitoring of inflammatory response.

Exogenous Intake of Fluorides in Caries Prevention: Benefits and Risks.

Caries incidence and prevalence have decreased significantly over the last few decades due to widespread use of fluoride. However, an increase in the prevalence of dental fluorosis has been reported in both fluoridated and non-fluoridated communities. Care must be taken to ensure that a balance between the optimal fluoride preventive effect at the individual and community level and minimal risk of dental fluorosis is maintained. This review describes the main sources of fluoride intake that have been identified: fluoridated drinking water, dietary fluoride supplement, and topical forms comprising toothpastes, rinses, gels and varnishes. The cited data were taken from meta-analytic studies and reports from Cochrane database systematic reviews up to December 2019. Efficiency, but safety, of topically applied fluorides in individual home care is dependent on the degree of compliance of individuals/parents and on the level of competence of providers of preventive counselling. The broad spectrum of these resources allows individualization of fluoride prevention based on risk analysis of caries attack and taking into consideration other preventive measures.

Evaluation of the risk factors of dental caries in children with very low birth weight and normal birth weight.

BACKGROUND: Health problems of premature infants can affect both general and oral health. The enamel defects, poor dietary and oral hygiene habits may predispose these children to dental caries. This study was conducted to assess the impact of very low birth weight and prematurity on caries risk in early childhood. METHODS: The study cohort consists of 189 of one year old infants. Anamnestic data were obtained from hospital records, feeding practice, bed-time sugar drinks and oral hygiene onset from questionnaires. Saliva samples of children and their mothers were collected for the detection of cariogenic pathogens. RESULTS: VLBW newborns had significantly shorter gestation age (29.6 vs. 38.8)) and lower mean birthweight (1124 g vs 3315 g) compared to NBW ones (p < 0.0001). Statistical significance has been found in the presence of early morbidity (p < 0.0001) and regular medication intake (p = 0.007). VLBW children got more frequently sweetened drinks during the day and night (p = 0.007). Regular oral hygiene practice was more frequent in full term group (p = 0.002). There was statistical difference in the presence of enamel hypoplasia in VLBW children (p = 0.033) but no statistical difference in the presence of hypomineralization (p = 0.0736) in comparison to NBW individuals. Proportional representation and count of S. mutans did not reveal statistical difference neither in both groups of children (p = 0.484) nor in both groups of mothers (p = 0.385). CONCLUSIONS: The study confirmed anamnestic and medical differences between both groups. The proportional representation and count of S. mutans did not reveal statistical difference neither in VLBW and NBW children, nor in their mothers.

Recurrent Aphthous Stomatitis in Children: A Practical Guideline for Paediatric Practitioners.

Recurrent aphthous stomatitis (RAS) is the most common chronic oral mucosal lesion affecting up to 25% of the population. The diagnosis is based on well-defined clinical characteristics, but the precise aetiology and pathogenesis remain unclear. The treatment of RAS should be based on the identification and control of possible predisposing factors. A wide range of topical medicaments is available as antiseptics, anti-inflammatory drugs and corticosteroids. The systemic treatment is indicated in patients with continuous and aggressive manifestation, which is extremely rare in children. The present article provides a review of the current concept and knowledge of the aetiology, pathogenesis, and management of RAS in the paediatric population.

Oral Manifestations of Nutritional Deficiencies: Single Centre Analysis.

INTRODUCTION: Oral manifestations of deficiency of iron, vitamin B12 and folic acid are thought to be common. Prevalence of these deficiencies among patients with compatible symptoms is not well known. The goal of this study was to summarize evidence from a dental practice of iron, vitamin B12 and folic acid deficiency in patients presenting with compatible oral manifestations. METHODS: 250 patients who presented with burning mouth syndrome, angular cheilitis, recurrent aphthous stomatitis, papillar atrophy of the tongue dorsum or mucosal erythema were identified. Patients underwent clinical examination, and the blood samples were taken. RESULTS: 250 patients (208 females; 42 males, mean age 44.1 years) with at least one corresponding symptom or sign were identified. The nutritional deficiency of one or more nutrients was found in 119 patients (47.6%). Seven times more females than males were noted to have one type of deficiency (104 females, 15 males). Iron deficiency as defined was diagnosed in 62 patients (24.8%), vitamin B12 or folic acid deficiency in 44 patients (17.6%) and both deficiencies (iron + vitamin B12/folic acid) in 13 patients (5.2%). The only predictive factor was gender and only for iron deficiency. The presence of more than one deficiency was noted in 10 patients (4.9%). CONCLUSION: The most commonly observed deficiency in dental practice over the course of 11 years was an iron deficiency in the female population. Age, diet and reported co-morbidities did not show statistically significant predictable value in recognizing these deficiencies.

Iron Deficiency as Cause of Dysphagia and Burning Mouth (Plummer-Vinson or Kelly-Patterson Syndrome): a Case Report.

The clinical presentation of iron deficiency can be very heterogeneous, including various oral and other mucosal problems. Here, in this case, we report the patient with burning mouth and dysphagia symptoms where iron deficiency was found to be the underlying cause after several months of investigations. This clinical syndrome is called Plummer-Vinson syndrome. It is sporadic with an incidence less than 0.1% of patients suffering from iron deficiency anemia.

Association between periodontal disease and preterm prelabour rupture of membranes.

OBJECTIVE: Periodontal disease is a possible contributing factor to preterm delivery. The aim of this study was to compare the periodontal status of women with preterm prelabour rupture of membranes (PPROM) and women with uncomplicated singleton pregnancies. PATIENTS AND METHODS: Seventy-eight women with PPROM at gestational ages between 24 + 0 and 36 + 6 weeks and 77 healthy women with uncomplicated pregnancies, matched for gestational age at sampling without preterm birth, were included in this study. All women underwent evaluation of periodontal and oral hygiene status. RESULTS: Women with PPROM had higher gingival and plaque indexes in crude analysis (gingival index: median 0.80 versus 0.20; p < 0.0001; plaque index: median 0.80 versus 0.10; p < 0.0001), even after adjustment for smoking status (p < 0.0001 and p < 0.0001). Mean clinical attachment loss (CAL) and probing pocket depth (PPD) values were higher in women with PPROM in the crude analysis (CAL: median 2.3 mm versus 1.8 mm; p < 0.0001; PPD: median 2.3 mm versus 1.8; p < 0.0001), as well as after adjustment for smoking status (p < 0.0001 and p < 0.0001). CONCLUSIONS: Pregnant women with PPROM residing in central Europe had worse periodontal status than women with uncomplicated pregnancies.

Levels of multiple proteins in gingival crevicular fluid and intra-amniotic complications in women with preterm prelabor rupture of membranes.

OBJECTIVE: The primary aim of this study was to identify the association between the local inflammatory response in gingival crevicular fluid measured by the levels of multiple proteins and maternal and intra-amniotic inflammatory responses measured by maternal serum C-reactive protein (CRP) and amniotic fluid interleukin (IL)-6 concentrations, respectively, in women with preterm prelabor rupture of membranes (PPROM). METHODS: A prospective study was performed in which 78 women with singleton pregnancies complicated by PPROM between 24 + 0 and 36 + 6 weeks of gestation were included. Transabdominal amniocenteses were performed at the time of admission. A bedside assessment of amniotic fluid IL-6 was performed. Maternal serum CRP concentration was also measured at the time of admission. Gingival crevicular fluid was collected from the pocket of the selected tooth (the tooth with the deepest pocket) using standard sterile paper strips within 72 h after admission. Twenty-six proteins in the gingival crevicular fluid were assessed by multiplex the Meso-Scale technology. RESULTS: No correlations between the levels of proteins in the gingival crevicular fluid and maternal serum CRP and amniotic fluid IL-6 concentrations were found, except for a weak positive correlation between granulocyte macrophage colony-stimulating factor and CRP. CONCLUSIONS: The local inflammatory response in the gingival crevicular fluid is not related to the maternal and intra-amniotic inflammatory responses in women with PPROM.

Periodontal disease and intra-amniotic complications in women with preterm prelabor rupture of membranes.

OBJECTIVE: Periodontal disease is frequently suggested as a possible causal factor for preterm delivery. The link between periodontal disease and preterm delivery is a possible translocation of periopathogenic bacteria to the placenta and amniotic fluid as well as a systemic response to this chronic inflammatory disease. However, there is a lack of information on whether there is an association between clinical periodontal status in women with preterm prelabor rupture of membranes (PPROM) and the presence of microbial invasion of the amniotic cavity (MIAC) and intra-amniotic inflammation (IAI). Therefore, the main aim of this study was to evaluate the incidence and severity of periodontal disease in women with PPROM. The secondary aim was to characterize an association between periodontal status and the presence of intra-amniotic PPROM complications (MIAC and/or IAI). MATERIALS AND METHODS: Seventy-eight women with PPROM at gestational ages between 24 + 0 and 36 + 6 weeks were included in this study. The samples of amniotic fluid were obtained at admission via transabdominal amniocentesis, and amniotic fluid interleukin (IL)-6 concentrations were determined using a point-of-care test. All women had a full-mouth recording to determine the periodontal and oral hygiene status. Probing pocket depth and clinical attachment loss were measured at four sites on each fully erupted tooth. RESULTS: In total, 45% (35/78) of women with PPROM had periodontal disease. Mild, moderate, and severe periodontal disease was present in 19% (15/78), 19% (15/78), and 6% (5/78) of women, respectively. The presence of MIAC and IAI was found in 28% (22/78) and 26% (20/78) of women, respectively. Periopathogenic bacteria (2 × Streptococcus intermedius and 1 × Fusobacterium nucleatum) was found in the amniotic fluid of 4% (3/78) of women. There were no differences in periodontal status between women with MIAC and/or IAI and women without these intra-amniotic complications. CONCLUSIONS: The presence of MIAC and IAI was not related to the periodontal status of women with PPROM.

Oral manifestation of sarcoidosis: A case report and review of the literature.

Sarcoidosis is a common multisystemic granulomatous disease characterized by the formation of nonnecrotizing epithelioid granulomas in various body organs. The most commonly affected organs in the head-and-neck region include salivary glands and cervical lymph nodes. Involvement of the oral mucosa is very rare. The purpose of this report is to present an atypical oral manifestation of sarcoidosis with ulcerations of the buccal mucosa in a 59-year-old female.

Dental and Oral Lesions in HIV-Positive Individuals in East Bohemia--Czech Republic, Single Centre Experience.

BACKGROUND: Human immunodeficiency virus (HIV) continues to be a serious health issue and one of the world most devastating epidemics. An estimated 1.5 million people died from AIDS-related illnesses in 2013, and an estimated 37 million people with AIDS have died worldwide since the epidemic has begun. HIV infection is known for its oral manifestations which causes discomfort and pain for infected individuals. The objective of this study was to document oral conditions of HIV positive patients and the pattern and frequency of oral and dental lesions. METHODS: All patients with confirmed HIV infection who were treated at the Department of Dentistry, University Hospital in Hradec Králové, were examined. RESULTS: During the study period, 29 HIV positive patients were examined and treated--19 men, 10 women, with mean age of 32.9 years (range 22-58 years). 72.41% patients received ART. In total, all patients underwent 186 visits. The most frequent treatments were associated with teeth and periodontal lesions (71.80%), oral mucosal lesions were diagnosed and treated only in 3.96% cases. CONCLUSION: Since the introduction of ART, the frequency of oral mucosal lesions is minimal in patients with HIV infection.

A retrospective study of 171 patients with oral lichen planus in the East Bohemia - Czech Republic - single center experience.

OBJECTIVES: Oral lichen planus is chronic inflammatory disease with a high prevalence in the population. This study describes the epidemiological and clinical characteristics of group of patients with oral lichen planus in the Czech Republic. MATERIAL AND METHODS: Data was taken from the medical records of 171 patients referred to the Oral Medicine Unit at the University Hospital in Hradec Králové with histologically confirmed clinical diagnosis of oral lichen planus in the period 2003 - 2013. The data were retrospectively reviewed. RESULTS: Of the 171 patients, 116 (67.8%) were women and 55 (32.2%) were men. The mean age was 55.2 ± 12.4 years (range of 85.0 - 20.9). The reticular form was the most frequent 93.6% (160 pts.), desquamative gingivitis was 12.9% (22). The buccal mucosa was the site most affected 89.5% (153 pts.). The lesions were asymptomatic in 52 patients (30.4%). Extraoral lesions were observed in 20.5% (35 pts.) of the patients, skin involvement was in 16.4% (28 pts.). Smokers were 29 patients. Local treatment used 116 (67.8%), only 6 patients used systemic short tome corticoid therapy. No evidence between OLP and malignant transformation was observed. CONCLUSIONS: This retrospective study show very similar profile and clinical features of the patients with OLP as in other studies. Key words:Oral lichen planus, clinical features, extraoral manifestation.