Gingival Crevicular Fluid Collection Research Articles

Effect of erbium, chromium-doped: yttrium, scandium, gallium, and garnet laser-assisted periodontal therapy using radial firing tip during early healing period: a randomized controlled split-mouth clinical trial

BackgroundThis study aimed to demonstrate the efficacy of erbium, chromium-doped:yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser-assisted nonsurgical periodontal therapy in periodontitis patients during 8 weeks of healing.MethodsA split-mouth, single-blinded, randomized controlled clinical trial was conducted on 12 patients diagnosed with stage III/IV periodontitis and had a minimum of two teeth with probing pocket depth (PPD) > 5 mm in at least two quadrants. Upon randomization, each quadrant was assigned for conventional scaling and root planing (SRP) procedure or laser-assisted therapy (SRP + laser) using radial firing tip (RFPT 5, Biolase). Clinical measurements and gingival crevicular fluid collection were performed for statistical analysis.ResultsIn the initial statistical analysis on the whole subject teeth, modified gingival index (MGI) reduction was greater in test group at 1(P = 0.0153), 4 (P = 0.0318), and 8 weeks (P = 0.0047) compared to the control in the same period. PPD reduction at 4 weeks in test group was -1.67 ± 0.59 showing significant difference compared to the control (-1.37 ± 0.63, P = 0.0253). When teeth with mean PPD ≥5 mm were sorted, MGI decrease was significantly greater in test group at 1 (P=0.003) and 8 week (P=0.0102) follow-ups. PPD reduction was also significantly greater in test group at 4 week period (-1.98 ± 0.55 vs -1.58 ± 0.56, test vs control, P=0.0224). ConclusionsEr,Cr:YSGG-assisted periodontal therapy is beneficial in MGI and PPD reductions during early healing period.

Association of Periodontitis and Atopic Dermatitis with the Levels of IL-13, IL-31, and TSLP in the Gingival Crevicular Fluid.

Emerging epidemiological evidence links atopic dermatitis (AD) and periodontitis, although the mechanisms remain unclear. Th2-derived cytokines are key in the development of both diseases, and different gingival crevicular fluid (GCF) profiles among healthy and diseased subjects have been previously reported. This case-control study examined the GCF levels of interleukins (IL)-13, IL-31, and thymic stromal lymphopoietin (TSLP) in 29 subjects with moderate-to-severe AD and 33 controls. All subjects underwent comprehensive clinical and oral evaluations, followed by GCF collection. GCF levels of IL-13, IL-31, and TSLP were assessed using a multiplex-bead immunoassay. Demographic and periodontal parameters were similar among groups (p > 0.05). The GCF levels of IL-31 and TSLP were higher in AD subjects compared to controls (p < 0.05), whereas no significant differences in the GCF levels of IL-13 were noticed (p = 0.377). Moderate-to-severe AD was positively associated with the GCF levels of IL-31 and TSLP, whereas severe periodontitis was negatively associated with IL-31 (p < 0.05). The GCF levels of IL-13 showed no significant associations with either condition (p = 0.689). There was no significant interaction between AD and periodontitis for IL-31 (p < 0.869). These results suggest that AD and periodontitis independently influence the GCF levels of IL-31 in opposing ways, whereas AD alone influences the levels of TSLP.

Orthodontically induced changes to the genetic profile in periodontal ligament tissue and cytokine release in gingival crevicular fluid – A pilot investigation

Background/purposeIt has been known that genetic factors influence orthodontic tooth movement, however, scientific research on humans is lacking. Therefore, this study aimed to investigate dynamic changes to the genetic profile in human periodontal ligament (PDL) tissue and cytokine release in gingival crevicular fluid (GCF) during the first 28 days of orthodontic treatment. Materials and methodsFifteen teeth from three patients were recruited. Full-mouth fixed appliances with extraction of four premolars and one maxillary third molar was planned for orthodontic treatment. GCF collection and tooth extraction were performed following force application for 0, 1, 3, 7, and 28 days. GCF was analyzed using multiplex immunoassay for 27 cytokines. PDL tissue was collected after extraction and submitted for RNA exome-sequencing using Illumina sequencing platform. Further analysis of differentially expressed genes (DEGs), gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and heatmaps were conducted. ResultsGCF cytokine levels varied among three patients; some patients exhibited a peak cytokine level on Day 0 whereas others did so on Days 1–3. In RNA exome sequencing data, GO and KEGG analyses showed that genes associated with sensory receptors were upregulated on Day 1, genes involved in bone remodeling were upregulated on Days 3 and 28, and genes related to osteoclast differentiation were upregulated on Day 7. ConclusionRNA sequencing data demonstrate that the specific types of genes are expressed at different time points, whereas the data on cytokine changes show a large variation in concentration levels and dynamic change patterns among the patients.

Effect of Cocoa Pod Husk Proanthocyanidins Extract on Cyclooxigenase-2 (COX-2) Expression in Gingival Crevicular Fluid of Periodontitis Rats

Inflammation of periodontal tissue due to Porphyromonas gingivalis which invades the gingival tissue due to the activity of virulence factors. This causes the host immune system changes and increases COX-2 production in periodontal tissue so resulting excessive tissue destruction. The aim of this study was to analyze the potency of proanthocyanidins extract of cacao pod husk to inhibit COX-2 expression in rat gingival sulcus fluid. 3 groups in this study, namely negative control, positive control, and treatment. In all groups, on the 0 day, gingival sulcus fluid was taken, then the rats were induced by Porphyromonas gingivalis once every 3 days for 2 weeks; then in the negative control group a placebo gel was applied and the treatment group was applied proanthocyanidins extract gel of cacao pod husk extract every day for 7, 14, and 28 days and serial gingival crevicular fluid collection was carried out on days 7, 14, and 28. The expression of COX-2 in the gingival crevicular fluid was observed using the ELISA method. The statistical test used was Anova. There were a significant difference in COX-2 expressions (p &lt;0.05). There was a decrease in the expression of COX-2 in rat gingival sulcus fluid induced by Porphyromonas gingivalis and given 10% cacao pod husk extract. Bioactive compounds in cocoa pod husk extract potential to reduce COX-2 expression in periodontitis rats. Article Details

BNT162b2 mRNA Vaccine–Induced Immune Response in Oral Fluids and Serum

The COVID-19 vaccine is currently being administered worldwide to address the ongoing pandemic. Although these vaccines have proven effective in preventing severe disease, the level of immunity required to prevent respiratory mucosal infection remains less well understood. Therefore, it is desirable to develop a noninvasive screening strategy such as oral fluid to monitor secreted antibodies longitudinally as potential surrogates of mucosal immunity. We evaluated the anti-spike protein antibodies in gingival crevicular fluid (GCF) and saliva and compared them to immune responses in the blood of 50 healthy health care workers following 2 doses of intramuscular Pfizer/BioNTech-BNT162b2 vaccine. The antibodies to SARS-CoV-2 spike and subdomain proteins (RBD, S1, S2, and NTD) were significantly higher in serum than oral fluids but showed a greater detection rate and higher median titres in GCF than saliva. For all tested SARS-CoV-2 antigens, IgG in GCF (as opposed to saliva) showed a more significant and stronger correlation with IgG in serum. Serum-neutralising antibodies (Nab) titres also displayed a significant and stronger correlation with anti-spike protein and their subdomains in GCF than saliva. Interestingly, the time post-second dose of vaccine and sex had a similar influence on IgG in serum and GCF. However, interferon (IFN)-γ-producing T-cell responses showed no association with SARS-Cov-2 IgG antibodies in serum, GCF, or saliva and neutralisation antibodies in serum. The correlation matrix of all measured parameters grouped serum and GCF IgG parameters separately from salivary IgG parameters indicating that GCF better represents the humoural response in serum than saliva. Within limitations, we propose that GCF could be a less invasive alternative to serum and more appropriate than saliva to detect antibody responses by current COVID-19 vaccines if the GCF collection procedure could be standardised. Further research is needed to investigate the suitability of GCF for community immune surveillance for vaccines.

Evaluation of injectable platelet-rich fibrin effect on the rate of canine retraction and alkaline phosphatase levels: An in-vivo study

With time, the treatment protocol has changed, and currently, there is a new school of treatment called accelerated orthodontics, wherein the goal is to shorten the time. In this study, a liquid formulation of platelet-rich fibrin such as injectable platelet-rich fibrin (i-PRF), was used, and its effect on the rate of canine retraction and the crevicular alkaline phosphatase (ALP) levels were studied. Thirteen patients were recruited for this study with a mean age of 20.6 ± 3.2 years. A split-mouth type of study design was used in which the maxillary arch of each subject was divided into an experimental and control group. I-PRF was injected in the labial and lingual attached gingiva of the canine in the experimental group. The gingival crevicular fluid collection was done from the distal aspect of the canine before canine retraction, 24 hours after retraction, and 28 days after retraction from both sites (ie, control and experimental sites). ALP activity was analyzed using a semiautomated analyzer, and the rate of canine retraction was measured on stone casts with the help of a digital vernier caliper. The individual canine retraction was 1.8-fold faster in the i-PRF group than in the control group. The ALP activity was significantly greater at 24 hrs and 28 days after retraction in the experimental group. These results suggest that i-PRF is an innovative, noninvasive approach to accelerating tooth movement. ALP activity in gingival crevicular fluid reflects the biological changes in the periodontium, and the steep increase in the activity indicates increased bone remodeling within the experimental group.

Comparative Evaluation of Gingival Crevicular Fluid Interleukin-17, 18 and 21 in Different Stages of Periodontal Health and Disease.

Background and Objectives: The elicitation of a host’s immune–inflammatory responses to overcome oral bacterial biofilm challenges is mediated by numerous cytokines. We explored the role of three such cytokines, viz. interleukin (IL)-17, 18 and 21, by measuring their levels in the gingival crevicular fluid (GCF) of Indian individuals with healthy gingiva, chronic gingivitis, or chronic periodontitis. Materials and Method: Ninety systemically healthy individuals were enrolled in the study on the basis of predefined criteria and were categorized into three groups of 30 participants each. Groups A, B and C were composed of a control group with healthy gingiva, subjects with chronic gingivitis and subjects with chronic periodontitis, respectively. The periodontal disease status was assessed on the basis of a subject’s gingival index, probing pocket depth, clinical attachment loss and radiographic evidence of bone loss. After the complete history-taking and identification of gingival sulcus/pocket depth areas for GCF collection, a sample was collected from each subject in all groups for an estimation of the cytokine levels using ELISA. Statistical analysis was performed using SPSS v 21.0. Intergroup comparisons were conducted using a post hoc Tukey’s test. A value of p < 0.05 was considered to be statistically significant. Results: The mean IL-17, 18 and 21 concentrations in pg/mL was the greatest for Group C (99.67 ± 18.85, 144.61 ± 20.83 and 69.67 ± 12.46, respectively), followed by Group B (19.27 ± 2.78, 22.27 ± 2.43 and 22.74 ± 1.43, respectively) and finally by Group A (healthy control; 11.56 ± 0.99, 17.94 ± 1.24 and 12.83 ± 1.21 respectively). A statistically significant difference in the mean concentrations of two interleukins (IL-17 and IL-18) was observed between Groups A and C and also between Groups B and C. A statistically significant difference in the mean concentrations of IL-21 was observed between Groups B and C. Conclusions: Within the limitations of the present study, the findings revealed that the GCF levels of IL-17, IL-18 and IL-21 rose and correlated well with the severity of the disease. Thus, these cytokines present in GCF have the potential to be considered as biomarkers for periodontal tissue destruction. IL-21 in particular appears to be a promising biomarker for differentiating between gingivitis and periodontitis.

Mapping of SARS-CoV-2 IgM and IgG in gingival crevicular fluid: Antibody dynamics and linkage to severity of COVID-19 in hospital inpatients

Mapping of SARS-CoV-2 IgM and IgG in gingival crevicular fluid: Antibody dynamics and linkage to severity of COVID-19 in hospital inpatients

Gingival crevicular fluid from pregnant women impairs trophoblast cell function and trophoblast‐neutrophil interaction

A strong association between periodontitis and higher susceptibility to pregnancy complications like preeclampsia has been reported although the mechanisms remain elusive. Trophoblast cells modulate the recruitment and functional shaping of maternal leukocytes at early stages to sustain an antiinflammatory microenvironment and fetal growth. Neutrophil activation with reactive oxygen species (ROS) release is associated with preeclampsia. Our aim was to study the effect of the gingival crevicular fluid (GCF) from pregnant women on trophoblast cell function and trophoblast-neutrophil interaction. Pregnant women at 16-20weeks of gestation (n=27) and non-pregnant women (n=8) as the control group were studied for gingivoperiodontal clinical score evaluation and GCF collection. Total bacteria and common periodontal pathogens were analyzed in GCF samples. The effect of each GCF sample was tested on first trimester trophoblast-derived cells to assess cell migration, cytokine expression and glucose uptake. Also, the effect of GCF on human peripheral neutrophil chemoattraction by trophoblast cells and ROS formation was assessed. Gingival crevicular fluid from pregnant women reduced trophoblast cell migration, increased proinflammatory marker expression and glucose uptake. A significant correlation between gingivoperiodontal score and trophoblast dysfunction was observed. Upon conditioning of trophoblast cells with GCF, only the GCF from pregnant women stimulated neutrophil chemoattraction. Similarly, GCF from pregnant but not from non-pregnant controls stimulated ROS formation in neutrophils. Gingival crevicular fluid from pregnant women is deleterious for first trimester trophoblast cell function. These effects could lead to placental homeostasis disruption underlying a pathogenic mechanism of pregnancy complications associated to periodontal disease.

Evaluation of salivary cytokines in patients with fixed orthodontic treatment

To evaluate the TNF-α and IL-1β levels in the gingival crevicular fluid (GCF) during orthodontic treatment. This study was carried out on 30 orthodontic patients. Prior to procedure, GCF was collected with a paper strip in gingival sulcus of maxillary right central incisor on the disto labial aspect for 60 seconds, followed by bonding and insertion of initial 14 NiTi archwire. GCF collection was repeated 24 hours, 7 days, and 30 days after bonding. Measurement of TNF-α and IL-1β levels was performed using Enzyme-Linked Immunosorbent Assay (ELISA). Data were evaluated statistically. There was raise in TNF-α and IL-1β concentration after treatment. There was no significant relationship between TNF-α and IL-1β concentrations subsequent to bonding in comparison to the baseline. There was no significant relationship in TNF-α and IL-1β level changes in the two age groups and among males and females. The present study indicated the raise in TNF-α and IL-1β concentration after treatment. There was no statically significant effect of gender and age on the concentration of TNF-α and IL-1β.

Collection, Storage and Protein Extraction Method of Gingival Crevicular Fluid for Proteomic Analysis

Gingival crevicular fluid (GCF) may reflect the events associated with orthodontic tooth movement. Attempts have been conducted to identify biomarkers reflecting optimum orthodontic force, unwanted sequallea (i.e. root resorption) and accelerated tooth movement. The aim of the present study is to find out a standardized GCF collection, storage and total protein extraction method from apparently healthy gingival sites with orthodontics that is compatible with further high-throughput proteomics. Eighteen patients who required extractions of both maxillary first premolars were recruited in this study. These teeth were randomly assigned to either heavy (225g) or light force (25g), and their site specific GCF was collected at baseline and after 1hr, 1day, 7days, 14days, 21days and 28days post force application. Periostrips were used for GCF collection and subsequent phosphate buffered saline (PBS) was used for immediate protein elution with centrifugal speed of 10000rpm for 5min and stored at -80°C. Protein concentration was estimated using Bradford colorimetric assay. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was carried out to resolve the purity of proteins in the collected samples and the method of collection was validated by western immuno-blotting of alpha amylase salivary enzyme. The current collection, storage and protein extraction protocol showed the best protein recovery and purity with validated collection free of salivary contamination. In conclusion, tiny GCF volume from healthy sites and evaporation issues of such promising non-invasive fluid motivate us to investigate a standardized protocol enabling optimal preservation of GCF sample and the currently followed protocol may serve as a reference for future proteomic studies searching for GCF biomarkers in diagnosing and monitoring orthodontic tooth movement.

Smoking effect on the interleukin-1β expression and periodontal status of periodontitis patients

Background: The smoking habit is a risk factor for periodontitis. Periodontitis is a multifactorial inflammatory disease associated with biofilm plaque dysbiosis and is characterized by the progressive deterioration of the periodontal tissue. Pro-inflammatory cytokines, such as interleukin-1β (IL-1β), can trigger inflammation of the tissue and become a stimulator to destroy it. Moreover, IL-1β will increase in response to inflammation. Objective: The aim of this study was to evaluate the differences in periodontal status and IL-1β gene expression between smoking and nonsmoking periodontitis subjects. Methods: This research was an analytic observational study with a cross-sectional design. The anthropometric demographic data included name, address, gender, age, plaque index examination, papillary bleeding index (PBI), calculus index, smoking status, and clinical photos to diagnose periodontitis. The collection of gingival crevicular fluid and the analysis of the IL-1β gene expression were conducted using real-time polymerase chain reaction. Results: There were significant differences in periodontal status for plaque index, calculus index, debris index, oral hygiene index, and PBI between smoking and nonsmoking subjects. The IL-1β gene expression was significantly higher (P = 0.047) in smoking subjects than in nonsmoking subjects, with mean ± standard deviation (11.34 ± 9.11) and (0.24 ± 0.42), respectively. Conclusion: This study revealed differences in periodontal status and the IL-1β gene expression between smok ing and nonsmoking periodontitis subjects. The periodontal rate was higher in periodontitis smokers compared to nonsmokers periodontitis. In this study, it was also found that the expression of the IL-1β gene was significantly higher in subjects with smoking periodontitis.

Effect of Corticosteroid Therapy on Tumor Necrosis Factor-Alpha during Orthodontic Tooth Movement with Self-Ligating Technique and Preadjusted Edgewise Technique and Alveolar Bone Loss in Partially Edentulous Patients

Background: The recent bracket systems are passive self-ligating orthodontic appliance systems, which have been shown to have superior load transfer toward the fibers of periodontal ligament than the preadjusted edgewise (PEA) orthodontic appliance systems. Corticosteroids have been shown to affect bone turnover, therefore, it is crucial to know if they affect orthodontic tooth movement and cortical bone resorption in partially edentulous patients. Hence, the aim of the present study was to analyze the effect of corticosteroid therapy on tumor necrosis factor (TNF)-alpha during orthodontic tooth movement with PEA appliance and self-ligating technique and alveolar bone loss in partially edentulous patients. Materials and Methods: Ninety-two participants were included in the study. They were divided randomly into four different groups. Group A: PEA appliance, Group B: self-ligating appliance, Group C: partially edentulous patients, and Group D: healthy controls. The test groups were administered injections of corticosteroids, namely triamcinolone acetonide, intramuscularly in the preparation of 1 mg/kg/day each day for 7 days. Control group research participants received a placebo injection. Gingival crevicular fluid (GCF) sampling was done with Gapadent #30 ISO Paper Points according to the Offenbacher et al. technique (1986). A human TNF-Quantikine enzyme-linked immunosorbent assay kit was used to quantify TNF-α contents in the samples. The GCF collection was performed using the same techniques at baseline, 1 h, 24 h, and 168 h after the bracket installation samplings. Results: Concentration of TNF-α decreased in all groups at 1 h and 7 days when compared with the baseline. When there was a comparison between the groups, then it was observed that TNF-α decreased in all three test groups compared to baseline. The decrease was more in the preadjusted appliance as compared with the self-ligating appliance. The decrease in the concentration of TNF-α was greater in the preadjusted appliance as compared with the self-ligating appliance showing that inflammatory reaction was more reduced in the preadjusted orthodontic appliance, and the tooth movement was affected more in such appliance after corticosteroid therapy. Conclusion: There was also a significant decrease in the concentration of TNF-α in partially edentulous subjects after corticosteroid therapy.

Tooth movement, orofacial pain, and leptin, interleukin-1β, and tumor necrosis factor-α levels in obese adolescents.

To evaluate tooth movement, orofacial pain, and leptin, interleukin (IL)-1β, and tumor necrosis factor (TNF)-α cytokine levels in the gingival crevicular fluid (GCF) during orthodontic treatment in obese adolescents. Participants included adolescent patients aged 12-18 years: group 1, obese (n = 30), and group 2, nonobese controls (n = 30). They were evaluated before (T0) and after 1 hour (T1), 24 hours (T2), and 1 week (T3) of fixed appliance bonding. Periodontal examination (T0), collection of GCF (T1, T2, T3), and evaluation of Little's irregularity index (T0, T3) were performed, and a visual analog scale was used to measure pain (T1, T2, T3). Evaluation of IL-1β, TNF-α, and leptin cytokines was performed using a Luminex assay. Mann-Whitney and t-tests were used for intergroup comparisons, and a generalized estimating equation and cluster analyses were used for comparisons among observation times (P < .05). The obese group had a higher prevalence of probing depth of ≥4 mm and bleeding on probing. Orthodontic tooth movement was similar in both groups. Peak of pain was at T2 in both groups and was higher in the obese patients. TNF-α showed a slight increase at T1, followed by a gradual decrease at T2 and T3 in both groups. The obese group had a higher concentration of IL-1β before and during orthodontic treatment. There was no difference in tooth movement between obese and control patients during the first week of orthodontic treatment. Obese adolescents had a greater subjective report of orofacial pain after 24 hours of orthodontic treatment and higher concentrations of IL-1β proinflammatory cytokine before and during tooth movement as compared with nonobese control adolescents.

A Comparative Study on the IL-8 Expression in Gingival Crevicular Fluid during Early Alignment Stage of Orthodontic Treatment in Adults and Adolescents

Bakground: Orthodontic tooth movement causes the release of various biomolecules such as interleukins. The aim of this study was to compare IL-8 expression in gingival crevicular fluid during early alignment stage of orthodontic treatment in adults and adolescents. Methods: The present study was done on 20 orthodontic patients, including 10 adolescents and 10 adults. Before bonding, gingival crevicular fluid (GCF) was collected with a paper strip in gingival sulcus of maxillary right central incisor on the distolabial aspect for 60 seconds, followed by bonding and insertion of initial 14 NiTi arch wire. GCF collection was repeated 24 hours, 7 days, and 28 days after bonding. The IL-8 levels was measured by the enzyme-linked immunosorbent assay (ELISA). Data were analyzed by SPSS version 21 using repeated measurement test. Statistically significant level was considered at P = 0.05. Results: In the present study, 14 patients (70%) were female and the rest were male. The mean age of the patients was 19.11 ± 6.23 years. The level of IL-8 at the baseline was higher than the other time periods, and on the first day after treatment, significantly decreased compared to the baseline. There was no statistically significant difference between age groups Conclusion: Theresults of the present study revealed that the level of IL-8 significantly decreased on the first day of orthodontic treatment, and then, increased. There was no statistically significant difference between gender and age and IL-8 levels during treatment. Further studies with larger sample sizes and different treatment methods are recommended.

Antimicrobial Photodynamic Therapy Using Curcumin Photosensitizer in Conjunction with Periodontal Open Flap Debridement. Pilot Clinical Study with Real-Time PCR Bacterial Assessment

Aim: To evaluate the effect of using antimicrobial photodynamic therapy (A-PDT) with curcumin in conjunction with open flap debridement (OFD) in the treatment of deep periodontal pockets. The primary objective was the clinical evaluation via assessment of the changes in clinical periodontal parameters. The secondary objective was the quantification of the pocket total bacterial load by Real-Time PCR. Subjects and methods: Group A included 7 patients who received OFD in conjunction with A-PDT using LED and curcumin paste (2%) as a photosensitizer. Group B included 7 patients who received OFD followed by application of curcumin paste (2%) but without photoactivation. Group C included 7 patients who received OFD alone. Clinical evaluation included plaque index (PI), gingival bleeding index (BI), probing depth (PD) and clinical attachment level (CAL). Microbiological evaluation involved collection of gingival crevicular fluid (GCF) samples for total bacterial load (TBL) analysis by real-time PCR Results: Regarding the percent change in PD and CAL, the greatest percent decrease value was recorded in group A while the least percent decrease was in group C. The difference between groups was statistically significant. Regarding percent change of TBL by time, the greatest percent decrease value was recorded in group A. Conclusion: Within the limitations of our study, the photoactivation of the curcumin used in conjunction with OFD may lead to greater PD reduction, greater CAL gain and greater TBL reduction when compared to OFD alone or OFD with curcumin without photoactivation.

Comparative evaluation of laser biostimulation as an adjunct to NSPT and ITS effects on AST levels in the management of chronic periodontitis: A randomized controlled trial

Aim: Clinical evaluation of aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) with and without laser biostimulation as an adjunctive to non-surgical periodontal therapy (NSPT). Materials and Methods: This is a split-mouth randomized trial. Ten patients (20 sites) were diagnosed with chronic periodontitis as per the American Academy of Periodontology (AAP) classification (1999), and radiographic evidence of horizontal bone loss was provided. Scaling and root planning (SRP) was performed in both the test groups. One of the test groups was treated with low-level laser therapy (LLLT) along with SRP and another with only SRP. GCF collection was done at baseline, during the third and sixth months by using microcapillary tubes for the estimation of AST. Quantification of AST was done by a Semi-automated Clinical Chemistry Analyzer. The data were subjected to statistical analysis by an independent t-test, Kruskal–Wallis test, Mann–Whitney U test, Wilcoxon signed-rank test, and post hoc pairwise comparison. Results: The results revealed a statistically significant reduction in pocket probing depth (PPD) (P < 0.001) and relative attachment gain (P < 0.001) at the test sites when compared from baseline till six months. Similarly, there was a statistically significant reduction in AST levels in GCF at the sites treated with laser biostimulation along with NSPT (P < 0.001). Conclusion: Laser biostimulation along with NSPT has shown propitious results as per clinical and biochemical marker analysis and evaluation.

ANALYSIS OF HERPES SIMPLEX VIRUS USING GINGIVAL CREVICULAR FLUID- A REVIEW

Oral Herpes is a viral infection caused by Herpes Simplex Virus. 80% of Oral Herpes is caused by HSV-1 which infects the skin and/or mucous membranes and produces painful sores or blisters on lips, gums, tongue, cheeks. The initial outbreak of virus strain can result in flulike symptoms, including fever, swollen lymph nodes and body aches. If not found in initial stages, it may result in systemic disorders like diabetes mellitus, cardiovascular or cerebrovascular diseases. The best solution to diagnose oral herpes is by analyzing Gingival Crevicular Fluid (GCF) through nested Polymerase Chain Reaction (PCR) technique. In periodontal disease pathogenesis, the inflammatory mediator levels present in gingival crevicular fluid represent relevant risk indicators for disease activity. Gingival crevicular fluid (GCF), a serum transudate or inflammatory exudate, can be collected from the gingival crevice surrounding the teeth. The fluid reflects the constituents of serum, the cellular response in the periodontium, and contributions from the gingival crevice. The amount of GCF is directly proportional to the severity of the inflammation. Now-a-days several methods for collection and estimation of GCF has been established. Hence, by analysing in this technique, the presence of HSV in the periodontium before the onset, at the time of periodontitis initiation, and periodically during its development can be diagnosed. KEYWORDS: Herpes virus, Gingival crevicular fluid, inflammation, periodontitis

Comparison of TNF-α and IL-1β Concentrations in Gingival Crevicular Fluid During Early Alignment Stage of Orthodontic Treatment in Adults and Adolescents

Objective: To compare the levels of TNF- α and IL-1 β in the gingival crevicular fluid (GCF) during early leveling stage of orthodontic treatment. Material and Methods: This study was carried out on 20 orthodontic patients, including 10 teenagers and 10 adults. Before bonding, GCF was collected with a paper strip in gingival sulcus of maxillary right central incisor on the disto labial aspect for 60 seconds, followed by bonding and insertion of initial 14 NiTi archwire. GCF collection was repeated 24 hours, 7 days, and 28 days after bonding. Measurement of TNF- α and IL-1 β levels was done utilizing Enzyme-Linked Immunosorbent Assay (ELISA). Data were analyzed in SPSS using repeated measurement test . Results: Concentration of TNF- α and IL-1 β increased after treatment. There was no significant relationship between TNF- α and IL-1 β concentrations subsequent to bonding in comparison to the baseline. There was no significant relationship in TNF- α and IL-1 β level changes in the two age groups and between males and females . Conclusion: Concentration of TNF- α and IL-1 β increased after treatment. Age and sex had no statically significant effect on the concentration of TNF- α and IL-1 β .

Estimation of macrophage inflammatory protein-1α (MIP-1 α) levels in serum and gingival crevicular fluid in periodontal health, disease, and after treatment—A clinico-biochemical study

Objectives: Previous studies have revealed high macrophage inflammatory protein-1α (MIP-1α) levels in the gingival crevicular fluid (GCF) and the serum of patients with chronic periodontitis. However, reports correlating the GCF and serum MIP-1α levels are inadequate. Therefore, the present study estimates the GCF and serum MIP-1α levels and its effects on periodontal health, disease, and after periodontal treatment.Materials and Methods: Periodontal examination and collection of GCF and serum was performed for 60 subjects categorized into four groups with 20 subjects in each group: Group I (healthy), group II (gingivitis), and group III (chronic periodontitis). Eight weeks after scaling and root planing, the GCF and serum were collected from 20 patients of group III, who were considered as group IV. MIP-1α levels were estimated using the enzyme-linked immunosorbent assay.Results: MIP-1α was detected in all samples. However, the levels reduced significantly in group IV (P < 0.05). Mean MIP-1α levels in GCF and serum were the highest in group III (1.481 ng/μL) and the lowest in group I (0.209 ng/μL), and those in groups II (0.685 ng/μL) and IV (0.276 ng/μL) appeared between those of groups I and III.Conclusions: The GCF and serum MIP-1α levels increased proportionally with the progression of periodontal disease (PD) and decreased after treatment. Because MIP-1α levels in the GCF and serum correlated positively with clinical parameters, MIP-1α may be considered a “novel biomarker” in PD progression. However, controlled longitudinal studies are required to confirm this possibility.