Electrolyzed Saline Targets Biofilm Periodontal Pathogens In Vitro.

Preventing the development and recurrence of periodontal diseases often includes antimicrobial mouthrinses to control the growth of the periodontal pathogens. Most antimicrobials are nonselective, targeting the symbiotic oral species as well as the dysbiosis-inducing ones. This affects the overall microbial composition and metabolic activity and consequently the host-microbe interactions, which can be detrimental (associated with inflammation) or beneficial (health-associated). Consequently, guiding the antimicrobial effect for modulating the microbial composition to a health-associated one should be considered. For such an approach, this study investigated electrolyzed saline as a novel rinse. Electrolyzed saline was prepared from sterile saline using a portable electrolysis device. Multispecies oral homeostatic and dysbiotic biofilms were grown on hydroxyapatite discs and rinsed daily with electrolyzed saline (EOS). Corresponding positive (NaOCl) and negative (phosphate-buffered saline) controls were included. After 3 rinses, biofilms were analyzed with viability quantitative polymerase chain reaction and scanning electron microscopy. Supernatants of rinsed biofilms were used for metabolic activity analysis (high-performance liquid chromatography) through measuring organic acid content. In addition, human oral keratinocytes (HOKs) were exposed to EOS to test biocompatibility (cytotoxicity and inflammation induction) and also to rinsed biofilms to assess their immunogenicity after rinsing. Rinsing the dysbiotic biofilms with EOS could reduce the counts of the pathobionts (>3 log10 Geq/mm2 reduction) and avert biofilm dysbiosis (≤1% pathobiont abundance), leading to the dominance of commensal species (≥99%), which altered both biofilm metabolism and interleukin 8 (IL-8) induction in HOKs. EOS had no harmful effects on homeostatic biofilms. The scanning electron micrographs confirmed the same. In addition, tested concentrations of EOS did not have any cytotoxic effects and did not induce IL-8 production in HOKs. EOS showed promising results for diverting dysbiosis in in vitro rinsed biofilms and controlling key periopathogens, with no toxic effects on commensal species or human cells. This novel rinsing should be considered for clinical applications.

Impact of low-level laser therapy as an adjunct to non-surgical periodontal treatment on the levels of tissue plasminogen activator and plasminogen activator inhibitor 1 in Stage 3-4, Grade C periodontitis patients: a split-mouth, randomized control study.

AIM: To investigate the effects of low-level laser therapy (LLLT) as an adjunct to non-surgical periodontal treatment (NSPT) on the plasminogen-activating system. MATERIALS AND METHODS: Stage 3-4 Grade C periodontitis and age-gender-matched healthy individuals participated in the split-mouth study (ClinicalTrials.gov identifier, NCT05233501). The study groups were Periodontitis/NSPT (Sham); Periodontitis/NSPT + LLLT (LLLT); Healthy (Control). Following NSPT, LLLT was applied on Days 0, 2 and 7. Clinical parameters were recorded at baseline and on Day 30. Gingival crevicular fluid (GCF) was collected at baseline, on days 7, 14, and 30; tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) levels were measured with ELISA. RESULTS: Clinical parameters, total GCF tPA (tPAt) and PAI-1 (PAI-1t) levels significantly reduced in LLLT and Sham groups (< 0.001). GCF tPAt levels in LLLT were significantly lower (< 0.05) than Sham on Day 7. GCF tPAt levels in periodontitis groups were significantly higher than the Control at baseline, on Days 7 and 14 (< 0.01). By Day 30, both groups decreased to control levels (> 0.05). GCF PAI-1t levels were significantly lower in LLLT than the Sham on day 30 (< 0.01), comparable to healthy controls (> 0.05). CONCLUSION: Adjunctive LLLT modulates the plasminogen activating system in severe periodontitis by altering GCF tPA and PAI-1 levels. CLINICAL RELEVANCE: LLLT as an adjunct to non-surgical periodontal treatment in patients with Stage 3-4 Grade C leads to reduced plasminogen activation.

The effect of low-level laser therapy as an adjunct to non-surgical periodontal treatment on gingival crevicular fluid levels of transforming growth factor-beta 1, tissue plasminogen activator and plasminogen activator inhibitor 1 in smoking and non-smoking chronic periodontitis patients: A split-mouth, randomized control study.

BACKGROUND AND OBJECTIVE: This study aimed to investigate the effects of low-level laser therapy (LLLT) as an adjunct to scaling and root planing (SRP) on smoking and non-smoking patients with chronic periodontitis. MATERIAL AND METHODS: The study was conducted using a split-mouth design with 30 patients with chronic periodontitis (15 smokers, 15 non-smokers) and 30 healthy individuals matched for age, sex and smoking status as controls. Groups were constituted as follows: Cp+SRP+Sham: non-smokers with chronic periodontitis treated with SRP; Cp+SRP+LLLT: non-smokers with chronic periodontitis treated with SRP+LLLT; SCp+SRP+Sham: smokers with chronic periodontitis treated with SRP; SCp+SRP+LLLT: smokers with chronic periodontitis treated with SRP+LLLT; C: control group comprised of periodontally healthy non-smokers; SC: control group comprised of periodontally healthy smokers. LLLT was first applied on the same day as SRP and again on days 2 and 7 after SRP treatment. Clinical parameters were recorded before non-surgical periodontal treatment (baseline) and on day 30. Gingival crevicular fluid samples were collected before periodontal treatment (baseline) and during follow-up visits on days 7, 14 and 30. Gingival crevicular fluid transforming growth factor (TGF)-ß1, tissue plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) levels were measured using enzyme-linked immunosorbent assay. RESULTS: All clinical parameters showed significant reductions between baseline and day 30 following SRP treatment in both the LLLT and sham groups (P<.001). No significant differences were observed between the LLLT and sham groups of either the smokers or non-smokers (P>.05). Gingival crevicular fluid PAI-1 levels decreased significantly in the SCp+SRP+sham and SCp+SRP+LLLT groups (P<.05), and gingival crevicular fluid tPA levels decreased significantly in the Cp+SRP+sham, Cp+SRP+LLLT and SCp+SRP+LLLT groups (P<.05). Gingival crevicular fluid TGF-ß1 levels decreased significantly in all treatment groups (P<.05). Although no significant differences were found between the gingival crevicular fluid PAI-1, tPA and TGF-ß1 levels of the LLLT versus sham groups (P>.05) at any of the time points measured, both LLLT groups showed significant reductions in tPA/PAI-1 ratios over time. CONCLUSION: Within the limits of this study, LLLT may be understood to play a role in the modulation of periodontal tissue tPA and PAI-1 gingival crevicular fluid levels, particularly in smoking patients with chronic periodontitis, and may thus be recommended as an adjunct to non-surgical periodontal treatment.

Ankaferd blood stopper enhances healing after osseous grafting in patients with intrabony periodontal defects.

BACKGROUND AND OBJECTIVE: The aim of this clinical study were to compare the clinical efficacy of ankaferd blood stopper (ABS) when used in combination with autogenous cortical bone graft (ACB) in the treatment of intrabony periodontal defects. MATERIAL AND METHODS: The study was planned as a split-mouth design. Fifteen patients with chronic periodontitis at 30 sites (six men, nine women; 42 ± 7 years) were included. Treatment sites had probing pocket depths (PPD) of ≥ 6 mm and osseous defect depths of ≥ 4 mm as radiographically assessed. Following the initial periodontal therapy, patients were randomly assigned to two treatments in contralateral areas of the dentition: ACB + ABS or ACB alone. At baseline and 6 mo after surgery, clinical parameters of plaque index, gingival index, PPD, clinical attachment level and gingival recession (GR) were recorded. The primary outcome variable was the change in clinical attachment level between baseline and 24 wk after surgery. Gingival crevicular fluid samples were collected immediately before surgery and at 2, 4, 6, 12 and 24 wk after the surgery. Gingival crevicular fluid volume was calculated and vascular endothelial growth factor levels in gingival crevicular fluid were measured. RESULTS: PPD decreased, clinical attachment level improved and gingival index decreased significantly in response to both modes of treatment (p < 0.05). Both treatment modalities resulted in a significant gain in radiographic bone levels compared to baseline (p < 0.05). Intergroup comparisons showed that there was a significantly higher gain in clinical attachment level in the ABS/ACB group compared to ACB group (p < 0.05) with significantly less GR (p < 0.05). Similarly, vascular endothelial growth factor concentration in gingival crevicular fluid was significantly higher in the ABS/ACB group at postoperative weeks 2 and 4 compared to the ACB group (p < 0.01). CONCLUSIONS: The findings suggest that ABS enhances the soft tissue healing during the periodontal defect fill by the ACB by stimulating angiogenesis and vascular endothelial cell function, prevents GR and thereby increases the clinical attachment gain.

Gingival crevicular fluid interleukin-8 and lipoxin A4 levels of smokers and nonsmokers with different periodontal status: a cross-sectional study.

BACKGROUND AND OBJECTIVE: Smoking is an important risk factor for periodontal disease and effects the pathogenesis of the disease. This study evaluated the impact of smoking on gingival crevicular fluid interleukin-8 (IL-8) and lipoxin A4 (LxA4 ) levels in patients with and without periodontal disease. MATERIAL AND METHODS: A total of 122 participants were grouped as follows: smokers with generalized aggressive periodontitis (S-GAgP, n = 15); smokers with chronic periodontitis (S-CP, n = 17); smokers with gingivitis (SG, n = 15); smokers classified as periodontally healthy (SH, n = 15); nonsmokers with generalized aggressive periodontitis (N-GAgP, n = 15); nonsmokers with chronic periodontitis (N-CP, n = 15); nonsmokers with gingivitis (NG, n = 15); and nonsmokers classified as periodontally healthy (NH, n = 15). Gingival index, plaque index, probing pocket depth and clinical attachment level were recorded. Gingival crevicular fluid IL-8 and LxA4 levels were analyzed by ELISA. RESULTS: Gingival crevicular fluid IL-8 levels varied among groups, as follows: S-GAgP>S-CP>SG>SH and N-GAgP>N-CP>NG>NH. The gingival crevicular fluid IL-8 levels were significantly higher in the S-GAgP group compared with the N-GAgP group and in the S-CP group compared with the N-CP group (p < 0.05); differences between the SG and NG and the SH and NH groups were not statistically significant (p > 0.05). Gingival crevicular fluid LxA4 levels also varied among groups, but in an inverse direction when compared with the IL-8 levels, as follows: S-GAgP 0.05). CONCLUSION: The study findings suggest that the observed increases in gingival crevicular fluid IL-8 levels and decreases in gingival crevicular fluid LxA4 levels reflect changes in immune and inflammatory responses that occur as a result of smoking.

Evaluation of gingival alterations in rats medicated with cyclosporine A, tacrolimus and sirolimus: a stereological study.

BACKGROUND AND OBJECTIVE: It has previously been shown that both cyclosporine A and tacrolimus cause gingival overgrowth in the rat. We proposed that sirolimus may play an important role in decreasing the severity of gingival overgrowth. Therefore, the aim of this study was to evaluate the gingival changes induced by immunosuppressants, in the presence and absence of sirolimus, using histopathology and stereological methods. MATERIAL AND METHODS: Thirty-six male Sprague-Dawley rats were distributed into six treatment groups, each containing six rats, as follows: (i) cyclosporine A for 8 wk; (ii) tacrolimus for 8 wk; (iii) sirolimus for 8 wk; (iv) cyclosporine A + sirolimus for 8 wk; (v) tacrolimus + sirolimus for 8 wk; and (vi) distilled water for 8 wk. Histomorphometric analyses included measurements of epithelial thickness and connective tissue width and height. Stereological analyses included measurements of volumetric densities of fibroblasts (Vf ), collagen fibers (Vcf ) and blood vessels (Vbv ). RESULTS: Connective tissue width and height were significantly increased in cyclosporine A, tacrolimus and cyclosporine A + sirolimus groups compared with the control group (p < 0.05), and epithelial thickness was significantly increased in the cyclosporine A group and tacrolimus group compared with the control group (p < 0.05). Vf was significantly increased in the cyclosporine A group and the tacrolimus group compared with the control group (p < 0.05), whereas Vcf and Vbv were significantly increased in the cyclosporine A, tacrolimus and cyclosporine A + sirolimus groups compared with the control group (p < 0.05). CONCLUSION: The results of the study suggest that sirolimus seems not to be associated with gingival overgrowth, and combined usage of sirolimus and immunosuppressants decreases the severity of gingival overgrowth.

Vascular endothelial cadherin and vascular endothelial growth factor in periodontitis and smoking.

OBJECTIVE: This study investigated the vascularization in periodontal disease process via revealing: (i) vascular endothelial cadherin (VE-cadherin) and vascular endothelial growth factor (VEGF) productions in periodontitis and (ii) the impact of smoking on this phenomenon. MATERIALS AND METHODS: Fifteen smokers and 15 non-smokers with/without periodontitis were allocated by split-mouth randomization regarding their smoking and periodontal statuses. The teeth with periodontitis in smokers (group 1), without periodontitis in smokers (group 2), with periodontitis in non-smokers (group 3), and without periodontitis in non-smokers (group 4) constituted the study groups. Gingival crevicular fluid (GCF) levels of VE-cadherin and VEGF were determined by ELISA to evaluate their profiles in the groups. RESULTS: There were increased VE-cadherin levels in groups 1 and 3 compared with groups 2 and 4 (P < 0.05). Group 2 demonstrated higher VE-cadherin level than group 4 (P < 0.05). Increased VEGF was noted in groups 1 and 3 compared with groups 2 and 4 (P < 0.05) with similar levels between groups 1 and 3 and groups 2 and 4 (P > 0.05). There were no correlations between the VE-cadherin and VEGF levels in all groups (P > 0.05). CONCLUSION: The results suggest that VE-cadherin and VEGF may increase in periodontitis, and smoking may uniquely cause VE-cadherin production in GCF.

The role of phosphatase and tensin homolog in drug-induced gingival overgrowth.

BACKGROUND AND OBJECTIVE: We proposed that phosphatase and tensin homolog (PTEN) might be one of the signaling proteins that alter the balance between cell growth and cell death in drug-induced gingival overgrowth. The aim of this study was to investigate the expression of PTEN in subjects using cyclosporine A and to analyze the relationship between PTEN and cell proliferation marker, proliferating cell nuclear antigen (PCNA), in cyclosporine A-induced gingival overgrowth. MATERIAL AND METHODS: In total, samples from 36 subjects, i.e. 24 cyclosporine A-mediated renal transplant patients with gingival overgrowth (n = 12) or without gingival overgrowth (n = 12) and 12 matched periodontally healthy subjects, were included in the study. PTEN and PCNA expressions in gingival tissues were analyzed using immunohistochemistry, PTEN expression was also analyzed by western blot. PTEN immunoreactivity was calculated with a histologic score (HSCORE) value and PCNA immunoreactivity was calculated with the PCNA-proliferative index. RESULTS: Phosphatase and tensin homolog HSCORE for the group with gingival overgrowth was found to be significantly lowest compared to the group without gingival overgrowth and the control group (p < 0.001) while the highest PTEN HSCORE was found in the control group. In addition, the PTEN HSCORE for the group without gingival overgrowth was significantly lower compared to controls (p < 0.001). The highest PCNA-proliferative index score was observed in the group with gingival overgrowth while the lowest score was observed in the control group (p < 0.001). The immunoblot signal for PTEN was significantly decreased in the group with gingival overgrowth compared to the group without gingival overgrowth and the control group (p < 0.001). Western blot results were different from immunohistochemistry and revealed there was no significant difference between the without gingival overgrowth and the control group (p > 0.05). CONCLUSION: Our results showing decreased PTEN levels in patients with gingival overgrowth supported with increased PCNA expression suggested that PTEN might take part in the imbalance between cell proliferation and death in drug-induced gingival overgrowth.

Activity of platelet activating factor acetylhydrolase following phase I periodontal therapy.

OBJECTIVE: Elevated levels of platelet activating factor (PAF), a potent inflammatory mediator in periodontal disease and decreased PAF levels following periodontal surgical therapy have been previously detected in gingival tissues and gingival crevicular fluid (GCF). Platelet activating factor acetylhydrolase (PAF-AH) is a calcium-independent phospholipase A2 that catalyses the hydrolysis of PAF, thereby inactivating this mediator The hypothesis, a relationship between activity of PAF-AH and healing following periodontal therapy, was tested by detecting activity of PAF-AH in GCF samples collected from sites that had undergone phase I periodontal therapy with generalized chronic periodontitis. METHODS: Twenty patients with generalized chronic periodontitis were divided into two groups (n = 10): group 1 with probing pocket depth (PPD) 4-5 mm and group 2 with PPD > or = 6-8 mm. Clinical parameters were recorded and GCF was sampled before phase I periodontal therapy and at the 2nd, 7th, 14th, 21st and 28th day follow-up evaluation visits. Activity of PAF-AH in GCF was analysed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Probing pocket depth at the 21st and 28th day in group 1, and PPD at the 14th, 21st and 28th day in group 2 were significantly decreased when compared to the baseline values (p < 0.001). Activity of PAF-AH (micromol/ml) was significantly decreased at the 7th, 14th, 21st and 28th day following phase I periodontal therapy in both groups 1 and 2 compared to the baseline values (p < 0.05). CONCLUSION: Platelet activating factor acetylhydrolase is detectable in GCF by ELISA and showed a continuous decrease following phase I periodontal therapy. Changes in the PAF-AH activity would be a progressive marker of periodontal healing to evaluate the success of periodontal therapies.

Activity of platelet activating factor acetylhydrolase following phase I periodontal therapy / Actividad de la acetilhidrolasa del factor activador de las plaquetas tras la fase I de la terapia periodontal

OBJECTIVE: Elevated levels of platelet activating factor (PAF), a potent inflammatory mediator, in perio-dontal disease and decreased PAF levels following periodontal surgical therapy have been previously detected in gingival tissues and gingival crevicular fluid (GCF). Platelet activating factor acetylhydrolase (PAF-AH) is a calcium-independent phospholipase A2 that catalyses the hydrolysis of PAF, thereby inactivating this mediator. The hypothesis, a relationship between activity of PAF-AH and healing following periodontal therapy, was tested by detecting activity of PAF-AH in GCF samples collected from sites that had undergone phase I periodontal therapy with generalized chronic periodontitis. METHODS: Twenty patients with generalized chronic periodontitis were divided into two groups (n = 10): group 1 with probing pocket depth (PPD) 4-5 mm and group 2 with PPD > 6-8 mm. Clinical parameters were recorded and GCF was sampled before phase I periodontal therapy and at the 2nd, 7th, 14th, 21st and 28th day follow-up evaluation visits. Activity of PAF-AH in GCF was analysed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Probing pocket depth at the 21st and 28th day in group 1, and PPD at the 14th, 21st and 28th day in group 2 were significantly decreased when compared to the baseline values (p < 0.001). Activity of PAF-AH (µmol/ml) was significantly decreased at the 7th, 14th, 21st and 28th day following phase I periodontal therapy in both groups 1 and 2 compared to the baseline values (p < 0.05). CONCLUSION: Platelet activating factor acetylhydrolase is detectable in GCF by ELISA and showed a continuous decrease following phase I periodontal therapy. Changes in the PAF-AH activity would be a progressive marker of periodontal healing to evaluate the success of periodontal therapies.

OBJETIVO: Niveles elevados del factor activador de las plaquetas (PAF) - un potente mediador inflamatorio en la enfermedad periodontal - y niveles disminuidos de PAF tras la terapia quirúrgica periodontal, han sido detectados previamente en los tejidos gingivales y el fluido crevicular gingival (FCG). La acetilhidrolasa del factor activador de las plaquetas(PAF-AH) es una fosfolipasa A2 independiente del calcio, que cataliza la hidrólisis de PAF, inactivando así este mediador. La hipótesis - la existencia de una relación entre la actividad de PAF-AH y la curación tras la terapia periodontal - fue sometida a comprobación mediante la detección de la actividad de PAF-AH en muestras de FCG recogidas de sitios que pasaron por la fase I de la terapia periodontal por periodontitis crónica generalizada. MÉTODOS: Veinte pacientes con periodontitis crónica generalizada fueron divididos en dos grupos (n = 10): grupo 1 con una profundidad de bolsa al sondeo (PPD) de 4-5 mm, y grupo 2 con PPD = 6-8 mm. Se registraron los parámetros clínicos, y se obtuvieron muestras de FCG antes de la fase I de la terapia periodontal, y en las visitas de evaluación de seguimiento los días 2, 7, 14, 21 y 28. La actividad de PAF-AH en FCG se analizó mediante ensayo por inmunoabsorción ligada a enzimas (ELISA). RESULTADOS: La profundidad de bolsa al sondeo los días 21 y 28 en el grupo 1, y PPD los días 14, 21 y 28 en el grupo 2 se vieron disminuidas significativamente cuando se les comparó con los valores iniciales (p < 0.001). La actividad de PAF-AH (µmol/ml) disminuyó significativamente los días 7, 14, 21 y 28 tras la fase I de la terapia periodontal en ambos grupos 1 y 2 en comparación con los valores al inicio del estudio (p< 0.05). CONCLUSIÓN: La acetilhidrolasa del factor activador de las plaquetases detectable en FCG mediante ELISA, y mostró una disminución continua tras la fase I de la terapia periodontal. Los cambios en la actividad de la PAF-AH sería un marcador progresivo de la curación periodontal para evaluar el éxito de las terapias periodontales.