Photobiomodulation with 808-nm diode laser enhances gingival wound healing by promoting migration of human gingival mesenchymal stem cells via ROS/JNK/NF-κB/MMP-1 pathway.

Photobiomodulation (PBM) has been shown to improve wound healing by promoting mesenchymal stem cell migration and proliferation. However, it remains unknown whether an 808-nm diode laser can influence human gingival mesenchymal stem cells (HGMSCs), and which dose this works well. In the present study, it was found that PBM could promote the migration of HGMSCs but not the proliferation. Furthermore, PBM could activate mitochondrial ROS, which could elevate the phosphorylation levels of JNK and IKB in HGMSCs, and further activate NF-κB as the nuclear translocation of p65 is elevated. Taken together, these present results indicate that PBM might promote cell migration via the ROS/JNK/NF-κB pathway.

Evaluation of Light-Emitting Diodes' Effects on the Expression Level of P53 and EGFR in the Gingival Tissues of Albino Rats.

Background and objectives: The light-curing unit is considered an essential piece of equipment in every dental office. This study was conducted to evaluate the effect of Light-Emitting Diodes (LEDs) by the light cure (LC) device on gingival tissues of albino rats histologically and by regarding the expression of P53 and epidermal growth factor receptor (EGFR). Materials and methods: Gingival tissues of the rats were exposed to LEDs for 30 s with an interval of 30 s for periods of 2 and 5 min and were examined after two and four weeks of light exposure. After the set time, histological sections were studied and the P53 and EGFR expressions were evaluated immunohistochemically and by molecular methods. Results: Mild hyperplasia and mild inflammatory response were detected in higher rates after two weeks of exposure when compared to 4 weeks postexposure. Whereas fibrosis was found at a higher rate after four weeks than that found after two weeks postexposure, parakeratosis was seen only in the group that was exposed for 5 min to LC and when biopsies were taken after 2 weeks. We found that the immunohistochemical expression of P53 was not changed. Similarly, the alteration of EGFR expression was statistically nonsignificant (p > 0.05) when compared to the control group. The data obtained from the qRT-PCR reaction was analyzed using the comparative CT (2-ΔΔCT) method. Statistically, there was no significant difference in the expression of EGER and P53 gene transcripts. Conclusions: LED causes no serious alteration in P53 and EGFR expression, and only trivial histopathological changes occurred, most of which recovered after a 4-week interval.

Effectiveness of laser adjunctive therapy for surgical treatment of gingival recession with flap graft techniques: a systematic review and meta-analysis.

Various flap graft techniques in the treatment of gingival recession have already been reported in the literatures for root coverage. Laser therapy has effects of ablative, hemostatic, and decontamination. Therefore, we performed a meta-analysis of randomized controlled trials (RCTs) to compare the efficacy of flap surgery combined with laser with surgery alone for treating gingival recession. The studies were searched from PubMed, Embase, Web of science, and the Cochrane Central Register of Controlled Trials by two reviewers up to August 2017. The quality of RCTs was assessed by Cochrane Handbook. Data were extracted from studies and analyzed by Review Manager 5.3. 95% confidence interval (CI) and risk ratio (RR) were calculated for dichotomous data. Seven RCTs with 173 patients and 296 teeth were included in the meta-analysis. We found no statistically significant differences between two groups in GRD (gingival recession depth) (P = 0.21), GRW (gingival recession width) (P = 0.92), RES (root esthetic score) (P = 0.21), and CRC (complete root coverage) (P = 0.09). Statistically significant differences were found between two groups in the WKT (width of keratinized tissue) (P < 0.0001) and 1-year follow-up of PD (probing depth) (P = 0.03) and CAL (clinical attachment level) (P < 0.00001). The meta-analysis found that surgery with laser therapy provided clinical advantages in terms of WKT and 1-year follow-up of PD and CAL. However, flap graft associated with laser did not offer additional benefit to root coverage and esthetics in treating gingival recession. More long-term studies are required to assess these parameters.

Comparison of gingival depigmentation with Er,Cr:YSGG laser and surgical stripping, a 12-month follow-up.

Gingival melanin hyperpigmentation is an esthetic concern for many individuals. In this study, we compared the standard surgical removal method with two different Er,Cr:YSGG laser settings in order to find the best treatment method. In 33 dental arches, the following three treatment groups were comparatively evaluated: (1) surgical stripping, (2) removal with laser setting 1 (4.5 W, 50 Hz, 100% water, 80% air, 60 µs, 800 µm Tip; MZ8), and (3) laser setting 2 (2.5 W, 50 Hz, 20% water, 40% air, 700 µs, 800 µm Tip; MZ8). We comparatively evaluated pain, patient satisfaction and wound healing, treatment time, and the amount of bleeding. Re-pigmentation was evaluated after 1 and 12 months by Hedin and Dummet pigmentation scores. Laser setting 1 had the best results regarding pain and patient satisfaction, although not statistically significant (P > 0.05). Wound healing results were better using lasers compared to surgical stripping (P < 0.05). Laser setting 1 was a faster procedure with mild amounts of bleeding. The least amount of bleeding was seen with laser setting 2. After 1 month, only two cases of the laser setting 2-treated areas showed an isolated pigmented area in the papilla; at 12 months, the mean Hedin indexes were still less than 2 and mean Dummett index less than 1 in all treatment techniques, with the lowest scores seen in the laser setting 1 sites. Based on our results, Er,Cr:YSGG laser can be more convenient for gingival depigmentation compared to surgical blade. Although not statistically significant, laser setting 1 with shorter pulse duration and higher water spray showed better overall results. However, laser setting 2, with longer pulse duration and less water spray, resulted in better coagulative effects and can be used to control bleeding wherever necessary in clinical practice.

Effects of high power-pulsed Nd:YAG laser irradiation on the release of transforming growth factor-beta (TGF-ß) and vascular endothelial growth factor (VEGF) from human gingival fibroblasts.

The purpose of this study was to investigate the effect of different high-power energy settings of a neodymium:yttrium-aluminum-garnet (Nd:YAG) laser (1064 nm) on cell viability of human gingival fibroblasts (GFs) and release of transforming growth factor-beta (TGF-ß) and vascular endothelial growth factor (VEGF) on these cells. GFs were isolated from human gingival connective tissues during the crown lengthening procedure. GFs were irradiated with different laser parameters as follows: group 1: 1 W (100 mJ, 10 Hz) 10 seconds; group 2: 1.5 W (150 mJ, 10 Hz) 10 seconds; group 3: 2 W (200 mJ, 10 Hz) 10 seconds; group 4: 1 W (100 mJ, 10 Hz) 20 seconds; group 5: 1.5 W (150 mJ, 10 Hz) 20 seconds; and group 6: 2 W (200 mJ, 10 Hz) 20 seconds. Cell viability/cell proliferation was analyzed with XTT (tetrazolium salt, cell proliferation kit) staining. The release levels of TGF-ß and VEGF were analyzed by the enzyme-linked immunosorbent assay. No significant differences were observed in the different laser irradiation groups compared to the control group in terms of cell viability (p > 0.05). The release of TGF-ß was not affected by different laser irradiation settings (p > 0.05). Only group 6 promoted significantly higher VEGF release from GFs in 24 hours compared to the control group (p ˂ 0.05). These findings suggest that high-power Nd:YAG laser is probably safe but has a very limited effect for wound healing.

Effects of laser biostimulation on the epithelial tissue for keratinized layer differentiation: an in vitro study.

Gingival augmentation techniques proposed in the international literature do not exclude a surgical component, which determines consequent post-surgical discomfort and results are not always predictable. In recent years, the introduction of laser biostimulation has led to a less invasive approach, particularly in the treatment of periodontally compromised patients, limiting the surgical phase to seriously compromised cases, with regeneration techniques for the restoration of a correct periodontal tissue anatomy. The aim of this in vitro study is to establish the validity of laser biostimulation in order to develop the epithelial keratinized layer of the tissue by stimulating fibroblasts-keratinocytes organotypic cultures and fibroblasts and keratinocytes mono-cultures. We created two groups (test and control), each one composed of 3 fibroblast cultures, 3 keratinocyte cultures and 3 organotypic cultures. We performed laser irradiation of test group with Wiser Doctor Smile Lambda, Flat Top Handpiece, at 50 J/cm2 of fluency with one application every 40 h for a total of 5 applications. Forty-eight h after the last laser application, we investigated the presence and amount of keratins 5 and 8 with citofluorymetric and western blotting analyses. Analyses showed an increase in keratin synthesis in test group cultures, showing a remarkable increase in production of keratin 8 in co-cultures test. Laser biostimulation can considerably enhance keratin synthesis when applied with high energy doses and repeated applications to keratinocytes-fibroblasts co-cultures.

Proliferation, migration, and expression of oral-mucosal-healing-related genes by oral fibroblasts receiving low-level laser therapy after inflammatory cytokines challenge.

BACKGROUND AND OBJECTIVES: Increased expression of inflammatory cytokines in the oral cavity has been related to the etiopathogenesis of oral mucositis and to delayed oral mucosal repair. Low-level laser therapy (LLLT) stimulates proliferation and migration of gingival fibroblasts, but the effects of specific inflammatory cytokines on oral mucosal cells and the modulation of these effects by LLLT have not been fully investigated. Therefore, this study investigated the effects of LLLT on oral fibroblasts after being challenged by oral-mucositis-related inflammatory cytokines. METHODS: Human gingival fibroblasts were seeded in plain culture medium (DMEM) containing 10% fetal bovine serum (FBS) for 24 hours. Then, cells were kept in contact with inflammatory cytokines (TNF-α, IL-1ß, IL-6, and IL-8) in serum-free DMEM for 24 hours. After this period, cells were subjected to LLLT with a diode laser device (LaserTABLE, InGaAsP, 780 nm, 25 mW) delivering energy doses from 0.5 to 3 J/cm2 . Irradiation was repeated for 3 consecutive days. Twenty-four hours after the last irradiation, cell migration (wound-healing and transwell migration assays), cell proliferation (BrdU), gene expression of COL-I and growth factors (real-time PCR), and synthesis of COL-I (Sirius Red assay) and VEGF (ELISA) were assessed. Data were subjected to two-way ANOVA and Tukey's tests or Kruskall-Walis and Mann-Whitney tests (P < 0.05). RESULTS: The inflammatory cytokines decreased the migration capacity of gingival fibroblasts. However, a statistically significant difference was observed only for IL-6, detected by transwell assay, where 30% less cells migrated through the pores (P < 0.05) and IL-8, with an increased wound area (116%; P < 0.05), detected by the wound healing method. Cell proliferation was not affected by contact with cytokines, while growth factors and COL-I expression (approximately 80%; P < 0.05), as well as VEGF synthesis (approximately 20%; P < 0.05), were decreased after contact to all tested cytokines. The opposite was seen for total collagen synthesis. LLLT promoted an acceleration of fibroblast migration (30%; P < 0.05) and proliferation (112%; P < 0.05) when delivering 0.5 J/cm2 to the cells previously in contact with the inflammatory cytokines. Gene expression of VEGF (approximately 30%; P < 0.05), and EGF (17%; P < 0.05), was stimulated by LLLT after contact with TNF-α and IL-6. CONCLUSION: LLLT can counteract the negative effects of high concentrations of inflammatory cytokines, especially IL-6 and IL-8 on gingival fibroblast functions directly related to the wound-healing process. Lasers Surg. Med. 48:1006-1014, 2016. © 2016 Wiley Periodicals, Inc.

The Influence of Shockwave Therapy on Orthodontic Tooth Movement Induced in the Rat.

Shockwave therapy is used in medicine due to its ability to stimulate healing processes. The application of orthodontic force evokes an inflammatory reaction resulting in tooth movement. Shockwave therapy might have an effect on both inflammatory and periodonal ligament cytokine profiles. Our aim was to evaluate the fluctuations of different inflammatory cytokines after orthodontic force induction with and without shockwave therapy. An orthodontic appliance was applied between the rats' molars and incisors. In conjunction with the commencement of orthodontic force, the rats were treated with a single episode of 1000 shock waves and the gingival crevicular fluid was collected for 3 days. The expression and concentration of different cytokines was evaluated by a commercial 4-multiplex fluorescent bead-based immunoassay. The level of all cytokines displayed a similar trend in both shockwave-treated and untreated groups; the concentration peaked on the first day and declined thereafter. In all cases, however, the cytokine levels were smaller in the shockwave-treated than in untreated animals; a significant difference was found for sRANKL and borderline difference for IL-6 on Day 1. We conclude that shockwave therapy during the induction of orthodontic tooth movement influences the expression of inflammatory cytokines.

Assessment of Photodynamic Inactivation against Periodontal Bacteria Mediated by a Chitosan Hydrogel in a 3D Gingival Model.

Chitosan hydrogels containing hydroxypropyl methylcellulose (HPMC) and toluidine blue O were prepared and assessed for their mucoadhesive property and antimicrobial efficacy of photodynamic inactivation (PDI). Increased HPMC content in the hydrogels resulted in increased mucoadhesiveness. Furthermore, we developed a simple In Vitro 3D gingival model resembling the oral periodontal pocket to culture the biofilms of Staphylococcus aureus (S. aureus), Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), and Porphyromonas gingivalis (P. gingivalis). The PDI efficacy of chitosan hydrogel was examined against periodontal biofilms cultured in this 3D gingival model. We found that the PDI effectiveness was limited due to leaving some of the innermost bacteria alive at the non-illuminated site. Using this 3D gingival model, we further optimized PDI procedures with various adjustments of light energy and irradiation sites. The PDI efficacy of the chitosan hydrogel against periodontal biofilms can significantly improve via four sides of irradiation. In conclusion, this study not only showed the clinical applicability of this chitosan hydrogel but also the importance of the light irradiation pattern in performing PDI for periodontal disease.

Anti-inflammatory effect of 635 nm irradiations on in vitro direct/indirect irradiation model.

Low-level laser therapy (LLLT) has been promoted for its beneficial effects on tissue healing and pain relief. As during laser treatment it is possible to irradiate only a small area of the surface body or wound and, correspondingly, of a very small volume of the circulating blood, it is necessary to explain how its photomodification can lead to a wide spectrum of therapeutic effects. To establish the experimental model for indirect irradiation, irradiation with 635 nm was performed on immortalized human gingival fibroblasts (IGFs) in the presence of Porphyromonas gingivalis lipopolysaccharides (LPS). The irradiated medium was transferred to non-irradiated IGFs which were compared with direct irradiated IGFs. The protein expressions were assessed by Western blot, and prostaglandin E2 (PGE2 ) was measured using an enzyme-linked immunoassay. Reactive oxygen species (ROS) were measured by DCF-DA; cytokine profiles were assessed using a human inflammation antibody array. Cyclooxygenase-2 (COX-2) protein expression and PGE2 production were significantly increased in the LPS-treated group and decreased in both direct and indirect irradiated IGFs. Unlike direct irradiated IGFs, ROS level in indirect irradiated IGFs was decreased by time-dependent manners. There were significant differences of released granulocyte colony-stimulating factor (G-CSF), regulated on activated normal T-cell expressed and secreted (RANTES), and I-TAC level observed compared with direct and indirect irradiated IGFs. In addition, in the indirect irradiation group, phosphorylations of C-Raf and Erk1/2 increased significantly compared with the direct irradiation group. Thus, we suggest that not only direct exposure with 635 nm light, but also indirect exposure with 635 nm light can inhibit activation of pro-inflammatory mediators and may be clinically useful as an anti-inflammatory tool.

Impact of extracorporeal shockwave therapy on tooth mobility in adult orthodontic patients: a randomized single-center placebo-controlled clinical trial.

AIM: This RCT investigated the effect of non-invasive extracorporeal shockwaves on tooth mobility in orthodontic patients after active treatment. MATERIALS AND METHODS: Seventy-two adult patients were included in the study. Immediately after active orthodontic treatment, patients were assigned to a treatment or a placebo group based on block randomization. The orthodontic patients were required to be otherwise healthy. The region of interest was the anterior portion of the mandible. The treatment group received a single shockwave treatment with 1000 impulses while the placebo group was treated with an acoustic sham. Tooth mobility was evaluated over a period of 6 months using a Periotest and manual testing. Pocket probing depths, bleeding on probing and the irregularity index were also assessed. RESULTS: Tooth mobility reduced significantly over 6 months in both groups, but shockwaves achieved significantly more rapid reduction on manual testing. Probing depth was significantly reduced while the irregularity index remained stable. Bleeding on probing was significantly reduced in the treatment group. No anti-inflammatory effect could be derived due to possible initial group differences. CONCLUSIONS: The mobility of teeth aligned by orthodontic treatment reduces over time. Shockwave treatment appeared to reduce tooth mobility more rapidly.

Selective cytotoxic effects of low-power laser irradiation on human oral cancer cells.

BACKGROUND AND OBJECTIVES: Low-power laser irradiation (LPLI) is known to regulate cell proliferation and migration in clinical use. Recent studies have shown that LPLI induces cell death in some certain types of cancer cell lines. However, the cytotoxic selectivity of LPLI for cancer cells is not fully understood. The aim of this study was to compare the cytotoxic effects of LPLI in both human oral cancer OC2 cells and normal human gingival fibroblast (HGF) cells. MATERIALS AND METHODS: LPLI at 810 nm with an energy density from 10 to 60 J/cm(2) was used to irradiate human oral cancer OC2 cells and normal HGF cells. RESULTS: We found that LPLI significantly diminished cell viability of human oral cancer OC2 cells due to cell cycle arrest at the G1 phase and the induction of cell death but that it had no or little effects on cell cycle progression and death in normal HGF cells. Moreover, the production of reactive oxygen species (ROS) and the loss of mitochondrial membrane potential (MMP) were elevated in human oral cancer OC2 cells compared with the un-irradiated cells. In contrast, these effects remained unchanged in normal HGF cells after exposure to LPLI. LPLI also induced apoptosis in caspase-3 dependent manner in human oral cancer OC2 cells, a mode of action that could be mediated by ROS and mitochondrial damage. CONCLUSION: Our findings imply LPLI might be a potential therapy for oral cancers.

Gingival tissue healing following Er:YAG laser ablation compared to electrosurgery in rats.

The erbium-doped yttrium aluminum garnet (Er:YAG) laser is currently used for periodontal soft tissue management with favorable outcomes. However, the process of wound healing after Er:YAG laser (ErL) treatment has not been fully elucidated yet. The aim of this study was to investigate the gingival tissue healing after ErL ablation in comparison with that after electrosurgery (ElS). Gingival defects were created in 28 rats by ablation with ErL irradiation or ElS. The chronological changes in wound healing were evaluated using histological, histometrical, and immunohistochemical analyses. The ErL-ablated gingival tissue revealed much less thermal damage, compared to the ElS. In the ElS sites, the postoperative tissue destruction continued due to thermal damage, while in the ErL sites, tissue degradation was limited and the defects were re-epithelialized early. Heat shock protein (Hsp) 72/73 expression was detected abundantly remote from the wound in the ElS, whereas it was slightly observed in close proximity to the wound in the ErL sites. Hsp47 expression was observed in the entire connective tissue early in the wound healing and was found limited in the wound area later. This phenomenon proceeded faster in the ErL sites than in the ElS sites. Expression of proliferating cell nuclear antigen (PCNA) persisted in the epithelial tissue for a longer period in the ElS than that in the ErL. The ErL results in faster and more favorable gingival wound healing compared to the ElS, suggesting that the ErL is a safe and suitable tool for periodontal soft tissue management.

Efficacy of the ND:YAG laser therapy on EBV and HSV1 contamination in periodontal pockets.

AIM: The aim of this retrospective multicenter study was to verify the efficacy of Nd:YAG laser in the treatment of periodontal pockets infected by Epstein-Barr Virus (EBV) and Herpes Simplex Virus 1 (HSV1). METHODS: Subgingival plaque samples of 291 Italian periodontal patients were analyzed by Real Time PCR to evaluate the frequency of both viruses before and after Nd:YAG laser-assisted periodontal treatment. RESULTS: Before treatment, EBV and HSV1 were observed in 29.9% and in 3.8% of periodontal patients respectively, while co-infection with both viruses was detected in 1.7% of cases. Periodontal Nd:YAG laser treatment ("Periodontal Biological Laser-Assisted Therapy", PERIOBLAST) produced statistical significant benefits, especially in EBV periodontal infection: 78.2% of EBV positive patients became EBV-negative following treatment. CONCLUSIONS: Results of this preliminary study highlight that EBV is found in periodontal pockets more frequently than HSV1, supporting the theory of the potential role of EBV in the onset and progression of periodontal disease. Moreover, our data showed that Nd:YAG laser-assisted periodontal treatment (Perioblast) is also effective in case of viral infection, validating evidences that it represents a successful alternative approach to traditional periodontal protocols.

The combined effects of irradiation and herpes simplex virus type 1 infection on an immortal gingival cell line.

BACKGROUND: Oral mucosa is frequently exposed to Herpes simplex virus type 1 (HSV-1) infection and irradiation due to dental radiography. During radiotherapy for oral cancer, the surrounding clinically normal tissues are also irradiated. This prompted us to study the effects of HSV-1 infection and irradiation on viability and apoptosis of oral epithelial cells. METHODS: Immortal gingival keratinocyte (HMK) cells were infected with HSV-1 at a low multiplicity of infection (MOI) and irradiated with 2 Gy 24 hours post infection. The cells were then harvested at 24, 72 and 144 hours post irradiation for viability assays and qRT-PCR analyses for the apoptosis-related genes caspases 3, 8, and 9, bcl-2, NFκB1, and viral gene VP16. Mann-Whitney U-test was used for statistical calculations. RESULTS: Irradiation improved the cell viability at 144 hours post irradiation (P = 0.05), which was further improved by HSV-1 infection at MOI of 0.00001 (P = 0.05). Simultaneously, the combined effects of infection at MOI of 0.0001 and irradiation resulted in upregulation in NFκB1 (P = 0.05). The combined effects of irradiation and HSV infection also significantly downregulated the expression of caspases 3, 8, and 9 at 144 hours (P = 0.05) whereas caspase 3 and 8 significantly upregulated in non-irradiated, HSV-infected cells as compared to uninfected controls (P = 0.05). Infection with 0.0001 MOI downregulated bcl-2 in non-irradiated cells but was upregulated by 27% after irradiation when compared to non-irradiated infected cells (P = 0.05). Irradiation had no effect on HSV-1 shedding or HSV gene expression at 144 hours. CONCLUSIONS: HSV-1 infection may improve the viability of immortal cells after irradiation. The effect might be related to inhibition of apoptosis.

Irradiation by light-emitting diode light as an adjunct to facilitate healing of experimental periodontitis in vivo.

BACKGROUND AND OBJECTIVE: This study evaluated the biostimulatory effect of 660 nm light-emitting diode (LED) as an adjunct in the treatment of experimental periodontitis. MATERIAL AND METHODS: Ninety-six Sprague-Dawley rats underwent experimental periodontitis by placement of a silk ligature followed with or without additive Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) injection. Irradiation with LED light was performed at varying energy densities of 5, 10 and 15 J/cm2, 1 d after debridement and detoxification. Rats were killed at 3, 7 and 14 d after irradiation with LED light, and the effect of irradiation was evaluated by descriptive histology and quantitative measurements of periodontal bone loss, inflammatory infiltration and cellular proliferation. RESULTS: Reduction of inflammation, accelerated collagen deposition and realignment was noted following irradiation with LED light at densities of 10 and 15 J/cm2, and temporary reduction of periodontal bone loss, as well as bundle bone apposition, was noted at day 3 in rats treated with 10 J/cm2 light. The biomodulatory effect was stronger in sites treated with Pg-LPS injection. In sites without Pg-LPS injection, temporary reduction of inflammation was noted in all LED light-irradiated specimens at day 3. No significant change in cellular proliferation was noted in any LED light-treated group. CONCLUSIONS: LED light (660 nm) with an energy density of 10 J/cm2 appeared suitable as an adjunct modality for periodontitis by temporarily reducing inflammation, facilitating collagen realignment and bundle bone deposition. Future studies will aim to amplify the biostimulatory effect of LED light by adding a supplementary medium or repeated irradiation.

Comparison of different focusing fiber tips for improved oral diode laser surgery.

BACKGROUND AND OBJECTIVES: State of the art for use of the fiber guided diode laser in dental therapy is the application of bare fibers. A novel concept with delivery fiber and exchangeable fiber tips enables the use of tips with special and optimized geometries for various applications. The aim of this study is the comparison of different focusing fiber tips for enhanced cutting efficacy in oral surgery. MATERIAL AND METHODS: For this purpose various designs of tip geometry were investigated and optimized by ray tracing simulations. Two applicators, one with a sphere, and another one with a taper, were realized and tested on porcine gingiva (diode laser, 940 nm, 5 W/cw; 7 W/modulated). The cutting depth and quality were determined by light microscope. Histological sections of the cuts were prepared by a cryo-microtome and microscopically analyzed to determine the cut depths and thermal damage zones. RESULTS: The simulations show that, using a sphere as fiber tip, an intensity increase of up to a factor of 16.2 in air, and 13.2 in water compared to a bare 200 µm fiber can be achieved. Although offering high focusing factor in water, the cutting quality of the sphere was rather poor. This is probably caused by a derogation of the focusing quality due to contamination during cutting and light scattering. Much better results were achieved with conically shaped fiber tips. Compared to bare fibers they exhibit improved handling properties with no hooking, more regular and deeper cuts (5 W/cw: 2,393 ± 468 µm, compared to the cleaved bare fiber 5 W/cw: 711 ± 268 µm). The thermal damage zones of the cuts are comparable for the various tips and fibers. CONCLUSIONS: In conclusion the results of our study show that cutting quality and efficiency of diode laser on soft tissue can be significantly improved using conically shaped fiber tips.

Effect of laser on TNF-alpha expression in inflamed human gingival tissue.

This study sought to evaluate the effect of low-level laser treatment combined with scaling and root planing (SRP) on gingival tissue levels of TNF-alpha in subjects with periodontal disease. Eighty gingival papilla biopsy samples were obtained from 60 patients diagnosed with chronic advanced periodontitis; randomly assigned to three treatment groups (n = 20), as well as 20 subjects with no periodontal disease (group A). Group B received SRP on a single quadrant/day for four consecutive days. On day 5, all quadrants were rescaled. Groups C and D received the same treatment as group B plus laser application with the low-level diode laser (630-670 nm, 1.875 J/cm(2)) for five and ten consecutive days, respectively. Papilla biopsies were obtained from subjects and evaluated by ELISA for levels of TNF-alpha. The values in the control group were 5.2 ± 3.21 pg/mg and baseline values for the examined groups were 46.01 ± 16.69. Significantly decreased level of TNF-alpha for groups C and D was found after treatment, while group B demonstrated reduction of TNF-alpha of 31.34%. The results of this study show suppression of TNF-alpha in gingival tissue after low-level laser treatment as adjunct to SRP. Data may suggest beneficial anti-inflammatory effects of the laser treatment when used as adjunctive periodontal treatment.

Inflammatory cytokines are suppressed by light-emitting diode irradiation of P. gingivalis LPS-treated human gingival fibroblasts: inflammatory cytokine changes by LED irradiation.

Human gingival fibroblasts (hGFs) play an important role in the inflammatory reaction to lipopolysaccharide (LPS) from P. gingivalis, which infects periodontal connective tissue. In addition, although light-emitting diode (LED) irradiation has been reported to have biostimulatory effects, including anti-inflammatory activity, the pathological mechanisms of these effects are unclear. This study examined the effects of 635-nm irradiation of P. gingivalis LPS-treated human gingival fibroblasts on inflammatory cytokine profiles and the mitogen-activated protein kinase (MAPK) pathway, which is involved in cytokine production. Gingival fibroblasts treated or not treated with P. gingivalis LPS were irradiated with 635-nm LED light, and cytokine profiles in the supernatant were assessed using a human inflammation antibody array. Expression of cyclooxyginase-2 (COX-2) protein and phosphorylation of extracellular signal-regulated kinase (ERK 1/2), p38, and c-Jun-N-terminal kinase (JNK) were assessed by Western-blot analysis to determine the effects on the MAPK pathway, and prostaglandin E(2) (PGE(2)) in the supernatant was measured using an enzyme-linked immunoassay. COX-2 protein expression and PGE(2) production were significantly increased in the LPS-treated group and decreased by LED irradiation. LPS treatment of gingival fibroblasts led to the increased release of the pro-inflammatory-related cytokines interleukin-6 (IL-6) and IL-8, whereas LED irradiation inhibited their release. Analysis of MAPK signal transduction revealed a considerable decrease in p38 phosphorylation in response to 635-nm radiation either in the presence or absence of LPS. In addition, 635-nm LED irradiation significantly promoted JNK phosphorylation in the presence of LPS. LED irradiation can inhibit activation of pro-inflammatory cytokines, mediate the MAPK signaling pathway, and may be clinically useful as an anti-inflammatory tool.

Assesment of radiotherapy effects on the blood flow in gingiva and dental pulp - a laser Doppler flowmetry study.

OBJECTIVE: This study aims to determine and compare the dental pulp and gingival blood flow in patients referred for oropharyngeal radiotherapy (RT) at three different time points: before the start, immediately after, and six months following the completion of RT. The aim is also to evaluate the dependence of the pulp and gingival blood flow on the radiation dose. METHODOLOGY: A prospective study included 10 patients referred for intensity-modulated RT (IMRT) in the oropharyngeal region, with at least one intact tooth surrounded by a healthy gingiva. The dose received by each selected tooth and adjacent gingiva was determined according to the map of treatment planning and computer systems. The blood flow measurements were performed using the laser Doppler flowmetry (LDF) method. RESULTS: Comparing vascular flows at three different time points, the median blood flow in the dental pulp showed no statistically significant difference (p=0.325), contrary to gingiva (p=0.011). Immediately after RT completion, the gingival flow significantly increased compared to its starting point (p=0.012). The pulp flow correlated negatively with the radiation dose, whereas a strong correlation was noted 6 months following the RT completion. CONCLUSIONS: RT caused a significant acute gingival blood flow increase, followed by a long-term (over six months) tendency to return to the starting levels. The dental pulp blood flow is differently affected by higher radiation doses (over 50Gy) in comparison to lower doses (below 50Gy). During RT planning, considering the possibility of protecting the teeth localized near the Gross Tumor Volume as a sensitive organ is recommended.