Saliva Of Oral Lichen Planus Patients Research Articles

Dysbiosis of saliva microbiome in patients with oral lichen planus

BackgroundOral microbiota is not only important for maintaining oral health but also plays a role in various oral diseases. However, studies regarding microbiome changes in oral lichen planus (OLP) are very limited. To the best of our knowledge, there has been only two studies investigating salivary microbiome changes in OLP. Therefore, the purpose of this study was to identify the characteristic microbial profile in the saliva of OLP patients, with or without erosive lesions, and compare that with recurrent aphthous ulcer (RAU), a common oral immunological disorder that also shows multiple erosive/ulcerative lesions. Whole saliva samples were collected from 20 patients with OLP (erosive E, n = 10 and non-erosive NE, n = 10), 10 patients with RAU (U) and 10 healthy controls (C). DNA was extracted from the saliva samples, and the 16S rDNA gene V4 hypervariable region was analyzed using Illumina sequencing.ResultsWe obtained 4949 operational taxonomic units (OTUs) from the V4 region in all saliva samples. Community composition analysis showed a clear decreased relative abundance of genera Streptococcus and Sphingomonas in saliva from RAU patients when compared to the other three groups. Relative abundance of Lautropia and Gemella were higher in E group, whereas relative abundance of Haemophilus and Neisseria were higher in NE group when compared to C group. Abiotrophia and Oribacterium were higher in OLP (combining E and NE groups), while Eikenella and Aggregatibacter were lower when compared to C group. There was statistically significance in α-diversity between E and RAU groups(p < 0.05). Significant differences in β-diversity were detected in bacteria between E and C; NE and C; as well as E and NE groups. The LDA effect size algorithm identified the g_Haemophilus might be the potential biomarker in NE group.ConclusionsWe found that salivary microbiome in erosive OLP was significantly different from that found in RAU; and these changes may be related to the underlying disease process rather than presence of ulcerative/erosive lesions clinically. In addition, our findings in bacterial relative abundance in OLP were significantly different from the previously reported findings, which points to the need for further research in salivary microbiome of OLP.

Study on the Role of Salivary Flora and NF-κB Inflammatory Signal Pathway in Oral Lichen Planus.

Oral lichen planus (OLP) is an inflammatory disease. It is believed that infection and immune dysfunction play a key role in its pathogenesis, but the specific mechanism of action remains unclear. The 16s rRNA high-throughput sequencing technique was used to analyze the microbial flora structure in the saliva of OLP patients and healthy controls. The relative abundance of Derxia, Haemophilus, and Pseudomonas in the saliva of the OLP group was lower than that of the healthy control group, but there was no significant difference in the overall structure of the microbial population. In addition, we measured the protein expression levels of toll-like receptor 4 (TLR4) and nuclear factor-kappab p65 (NF-κB p65) in the tissues of OLP patients, and found that there was a significant increase and positive correlation between them (r = 0.907, P = 0.034). The expression levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the OLP group were consistent with those of NF-κB p65. Therefore, we believe that changes in the composition ratio of microbialflora break the original balance state of flora, promote the occurrence of immune inflammatory reaction, and then lead to the generation or aggravation of OLP disease. This discovery provides new ideas for further research on OLP initiation and immune regulation mechanism.

Cytokines, cortisol, and nitric oxide as salivary biomarkers in oral lichen planus: a systematic review

The etiopathogenesis of oral lichen planus (OLP) is still not fully elucidated, and it is believed that its development could involve a neuro-immune-endocrine profile. This systematic review investigated the relationship between cytokines, cortisol, and nitric oxide (NO) in the saliva of OLP patients. An electronic search was conducted in Pubmed/Medline, Scopus, LIVIVO, and Web of Science databases with no restriction of language to identify studies published up to December 2017. Data extraction was performed using the Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guidelines. A total of 140 articles were retrieved, and 32 articles fulfilled the inclusion criteria (cytokines = 17; cortisol = 9; NO = 6). The most studied cytokines in the saliva of OLP patients were interleukins IL-4, IL-6, IL-8, IFN-ү, and TNF-α, which were higher in OLP patients than in healthy controls (HC). Salivary cortisol was found to be higher in OLP than in HC in most (55.5%) of the selected studies, and all studies related to NO found higher levels of this marker in OLP than in HC. Despite controversial results, our review suggests that OLP patients have an increased inflammatory response, as indicated by the proinflammatory profile of salivary cytokines. In addition, we conclude that salivary cytokine and NO measurements may have significant diagnostic and prognostic potential for monitoring disease activity and therapeutic responses in OLP.

Putative salivary protein biomarkers for the diagnosis of oral lichen planus: a case-control study

BackgroundSalivary protein biomarkers for screening and diagnosis of oral lichen planus (OLP) are not well-defined. The objective of this study was to identify putative protein biomarkers for OLP using proteomic approaches.MethodsPooled unstimulated whole saliva was collected from five OLP patients and five healthy control participants. Saliva samples were then subjected to two-dimensional gel electrophoresis, followed by mass spectrometry to identify putative protein biomarkers. Subsequently, a subset of these putative biomarkers were validated in 24 OLP patients and 24 age-matched healthy control subjects, using an enzyme-linked immunosorbent assay (ELISA). Immunoblotting analyses were then performed in 3 pairs of age- and sex-matched OLP patients and healthy controls to confirm results from the ELISA study.ResultsThirty-one protein spots were identified, corresponding to 20 unique proteins. Notably, fibrinogen fragment D and complement component C3c exhibited increased expression in OLP patients, while cystatin SA exhibited decreased expression in OLP patients, compared with healthy control subjects. ELISA analyses indicated increased expression of fibrinogen fragment D and complement component C3c, and decreased expression of cystatin SA, in the saliva of OLP patients. Statistical differences in the expression of salivary complement C3c were observed between OLP patients and healthy control subjects. Immunoblotting analyses confirmed the results of our ELISA study.ConclusionComplement C3c, fibrinogen fragment D and cystatin SA may serve as salivary biomarkers for screening and/or diagnosis of OLP.

Diagnostic and prognostic relevance of salivary microRNA-21, -125a, -31 and -200a levels in patients with oral lichen planus - a short report.

Oral lichen planus (OLP), a relatively common chronic inflammatory disease of the oral mucosa, is considered to be a premalignant disorder of the oral cavity. Previously, several biomarkers have been tested for their diagnostic potential. Here, we aimed to investigate the diagnostic potential of four miRNAs, miR-21, -125a, -31 and -200a, known to be involved in oral squamous cell carcinoma (OSCC) development, in the saliva of OLP patients as also their putative relation to OSCC development in these patients. Saliva samples from 30 patients with OLP were collected, 15 of whom were diagnosed with dysplasia upon histopathologic examination. In addition, 15 saliva samples from patients with OSCC and 15 saliva samples from healthy donors were collected. After RNA extraction, the respective miRNA levels were assessed by quantitative RT-PCR. We found that the miR-21 levels were significantly increased in saliva samples derived from patients with OLP, dysplastic OLP and OSCC, compared to those from healthy controls (p = 0.012, p = 0.0017 and p < 0.0001, respectively). Conversely, significant decreases in miR-125a levels were found in the OLP, dysplastic OLP and OSCC samples, compared to those from healthy controls (p < 0.0014, p < 0.0001 and p < 0.0001, respectively). In addition, significant increases in miR-31 levels were found in samples derived from dysplastic OLP and OSCC patients, but not in those from nondysplastic OLP patients, compared to those in healthy controls (p = 0.01 and p = 0.004, respectively). Finally, we found that the miR-200a levels were significantly decreased only in samples derived from OSCC patients (p < 0.0001). From our data we conclude that increased miR-21 levels in conjunction with decreased miR-125a levels in saliva of OLP patients may be indicative for a poor prognosis. Conversely, we conclude that lack of significant alterations in miR-31 and miR-200a levels in saliva of OLP patients may be indicative for absence of malignant transformation.

Evaluation of oxidative stress markers in oral lichen planus.

Background:Lichen planus (LP) is a chronic, inflammatory disease that affects the skin, mucous membrane, scalp and nails that frequently involves the oral mucosa. Oxidative stress reflects an imbalance between the production of reactive oxygen species and the biological system's ability to readily detoxify the reactive intermediates or repair the resulting damage. It has been suggested that oxidative stress may play a role in the pathogenesis of LP.Aim and Objectives:To evaluate the role of oxidative parameters in the pathogenesis of oral LP, estimate the levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPx) and uric acid (UA) in saliva of oral LP patients and to compare the levels of SOD, MDA, GP and UA in oral LP patients with healthy controls.Methodology:In this cross-sectional study, 1.5 ml of fasting saliva sample was collected using passive drool method from the study group (30 patients diagnosed as having oral LP) and the control group (30 age-matched healthy volunteers). The unstimulated saliva was collected were analyzed by spectrophotometry. Statistical analysis was done to evaluate and compare the values between groups.Results:The mean values of SOD and MDA in saliva in the study group showed a significant increase in amount when compared with the control group whereas GPx showed a significant decrease in the study group. UA value showed an insignificant difference in the same comparison.Interpretation and Conclusion:Oxidative stress markers as MDA and SOD are elevated, and GPx is decreased in the saliva of oral LP patients.

Evaluation of salivary oxidate stress biomarkers, nitric oxide and C-reactive protein in patients with oral lichen planus and burning mouth syndrome.

The aim of this study was to evaluate oxidative stress factors and C-reactive protein in the saliva of patients with oral lichen planus (OLP) and burning mouth syndrome (BMS). This consecutive, cross-sectional study included 20 patients with OLP, 19 with burning mouth syndrome (BMS), and 31 control subjects. The oral cavity of each patient was examined and patients responded to a quality of life questionnaire (OHIP-14) and the xerostomia inventory. The following parameters were measured in whole non-stimulated saliva: trolox equivalent antioxidant capacity (TEAC); total antioxidant capacity (TAC); cupric reducing antioxidant capacity (CUPRAC); ferric reducing ability of plasma (FRAP); C-reactive protein (CRP); nitric oxide; nitrates; and nitrites. The OLP group presented statistically significant differences in reactive oxygen species (ROS) (29 600 cps) in comparison with the control group (39 679 cps) (P < 0.05). In the BMS group, ROS was 29 707 cps with significant difference in comparison with the control group (P < 0.05). Significantly higher salivary nitric oxide (145.7 μmol) and nitrite (141.0 μmol) levels were found in OLP patients in comparison with control group (P < 0.05). Increases in nitric oxide and C-reactive protein were found in the saliva of OLP patients in comparison with BMS and control patients. Further studies are required to confirm these findings.

Analysis of oral microbial community and Th17-associated cytokines in saliva of patients with oral lichen planus.

Oral lichen planus (OLP) is a chronic inflammatory disorder of oral mucosa of unknown cause. Microbial infection and dysimmunity appear to play important roles in its pathogenesis. In this study, differences in genetic profiling of salivary microbial communities in two subtypes of OLP and healthy controls were evaluated by means of PCR-denaturing gradient gel electrophoresis (DGGE). Additionally, ELISA was used to investigate the possible role of Th17 in lesion formation by detecting two related cytokines IL-17 and IL-23 in the saliva of OLP patients. When the DGGE profiles were analyzed, the bacterial populations were found to be significantly less rich in subjects with reticular and erosive OLP than in healthy controls. There was significantly less microbial diversity, as denoted by the Shannon index, in saliva samples from subjects with erosive OLP than in those from healthy controls. Cluster analysis and principal component analysis showed that the DGGE profiles formed distinctly group-specific clusters. Salivary concentrations of IL-17 in subjects with erosive OLP group were significantly higher than in those with reticular OLP and healthy controls. What's more, significantly positive correlations were observed between salivary IL-17 concentrations and disease clinical scores. Microbial richness and diversity was negatively correlated with salivary IL-17 concentrations. These results suggest there is significantly less salivary bacterial diversity and complexity in subjects with OLP han in healthy controls and that the shifted community composition is closely related to an immune cytokine, IL-17.

IFN-Gamma and IL-4 in Saliva of Patients with Oral Lichen Planus: A Study in an Ethnic Chinese Population

Interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) represent T helper 1 (Th1) and T helper 2 (Th2) cytokines involved in oral lichen planus (OLP), respectively. This study was to investigate the expression profile of IFN-gamma and IL-4 in saliva of OLP patients. Seventy-nine ethnic Chinese patients with OLP were recruited for this study, together with 41 age-sex-matched healthy volunteers served as control group. IFN-gamma and IL-4 levels in whole unstimulated saliva were screened by enzyme linked immunosorbent assay. OLP patient showed a low-level IFN-gamma but high-level IL-4 expression profile in saliva, with a lower ratio of salivary IFN-gamma/IL-4 compared to healthy controls. With regards to subtypes, salivary IL-4 level in erythematous/ulcerative group was significantly higher than that in reticular group. Imbalance of Th1/Th2 cytokines with Th2-predominant profile in saliva may be involved in OLP. Salivary IL-4 level may be a fine biomarker reflecting the severity of OLP.

Alteration of the Expression of CD4 Isoforms in Oral Epithelia and Saliva from Patients with Oral Lichen Planus

Oral lichen planus (OLP) is a chronic inflammatory mucosal disease that cell-mediated immunological mechanisms are involved in pathogenesis. The objective of this study was to investigate the expression of CD44 isoforms including CD44s, CD44v5, and CD44v6 in biopsy specimens and saliva from OLP patients. Thirty-one OLP patients and 30 healthy subjects were enrolled in this study. Immunohistochemical methods were used to detect the expression of CD44 isoforms in oral epithelia, and enzyme-linked immunosorbent assay (ELISA) was performed to measure levels of salivary CD44 isoforms. Our results demonstrated that expression of CD44v6 in oral epithelia from OLP patients was significantly decreased in comparison to controls (p = 0.021). Levels of salivary CD44s and CD44v5 from OLP patients were significantly higher than those from controls (p = 0.007 and p = 0.002, respectively). In summary, our findings provided additional evidence that the pathological stress, such as chronic inflammation, altered the expression of CD44 isoforms in oral epithelia and saliva of OLP patients.

Comparative analysis of whole saliva proteomes for the screening of biomarkers for oral lichen planus

To screen possible biomarkers in whole saliva from oral lichen planus (OLP) patients by proteomic techniques. Two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were applied to whole saliva of OLP patients and controls, and the differential protein components were analyzed with peptide mass fingerprinting and database searching. A total of 31 protein spots representing 14 proteins with at least two-fold difference in abundance between OLP and controls were identified. Among these, the expression of urinary prokallikrein was increased while short palate, lung and nasal epithelium carcinoma associated protein (PLUNC) was decreased in all samples of OLP. Urinary prokallikrein and PLUNC may be the new biomarkers which play a role in inflammation and immune response of OLP.