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Near-infrared (NIR) irradiation has shown potential to stimulate osteogenic differentiation, but the mechanisms are not fully understood. The study is to investigate the effects of NIR laser irradiation on osteoblastic differentiation. Human periodontal ligament stem cells (hPDLSCs) were cultured in osteogenic medium and exposed to 810 nm NIR laser at 0.5 J/cm 2 every 48 h. The transient receptor potential vanilloid (TRPV1) channel inhibitor capsazepine (CPZ) was used to evaluate the role of calcium influx. Osteogenic differentiation was assessed by proliferation (CCK-8), alkaline phosphatase (ALP) activity, mineralization (Alizarin Red), and expression of bone markers by PCR and Western blot over 2 weeks. Intracellular calcium was measured by Fluo-4M dye and flow cytometry. Results showed that NIR irradiation enhanced hPDLSC proliferation, ALP activity, mineralization, and bone marker expression, indicating increased osteogenic differentiation. These effects were inhibited by CPZ. NIR induced a transient rise in intracellular calcium peaking at 3 min, which was blocked by CPZ. In conclusion, this study demonstrates that NIR laser irradiation promotes osteogenic differentiation of PDLSCs through the activation of TRPV1 channels and subsequent calcium signaling. Further research is warranted to optimize the treatment parameters and elucidate the detailed signaling pathways involved, paving the way for the clinical application of NIR therapy in the treatment of bone disorders and periodontal disease.

Background: The present study aimed to investigate the effects of infrared diode laser irradiation on the proliferation and differentiation capacity of periodontal ligament stem cells (hPDLSCs), which are optimal cell sources for periodontal regeneration., Methods: hPDLSCs were isolated and characterized by flow cytometric analysis of mesenchymal stem cell markers, and their trilineage differentiation capacity was tested. hPDLSCs were then cultured and irradiated with infrared diode laser (970 nm) at a power of 200 mW and a fluence of 4 J/cm 2 for 3 s. MTT assay was performed to assess cellular proliferation. Cell cycle analysis was performed, and the impact of infrared diode laser irradiation on the stemness and osteogenic differentiation potential of hPDLSCs was evaluated via RT‒PCR., Results: Infrared diode laser application enhanced the stemness, viability, proliferation, and differentiation of PDLSCs. Stem cell markers (OCT4, SOX2, and NANOG) were significantly upregulated in hPDLSCs exposed to laser irradiation. There was significant overexpression of RUNX2, ALP, OPN, and OCN on day 14 after laser application., Conclusions: These findings provide valuable insights into the specific applications of infrared diode lasers to effectively regenerate periodontal tissues. The results can aid in the development of precise clinical protocols aimed at enhancing osseointegration and promoting tissue regeneration. Ultimately, the combination of infrared diode laser with hPDLSCs is promising for stimulating periodontal regeneration., (© 2024. The Author(s).)

This study assessed the impact of low-level laser irradiation on the viability and proliferation of human periodontal ligament stem cells (hPDLSCs) cultivated on polylactic acid (PLA) scaffolds. hPDLSCs were obtained, characterized, and grown on the surface of PLA films produced via the solvent casting technique. The study involved two groups: the control group, which was not exposed to radiation, and the laser group, which was irradiated with a diode laser (InGaAIP) with a power of 30 mW, a wavelength of 660 nm, and a single dose of 1 J/cm² emitted continuously. Cell viability was assessed 24 and 48 hours after irradiation using the Alamar blue and Live/Dead assays. Flow cytometry was used to assess cell cycle events, and scanning electron microscopy (SEM) was used to evaluate the interaction between cells and the biomaterial. The results revealed a statistically significant increase in cell metabolic activity in the laser group compared with the control group at 24 hours (p <0.05) and 48 hours (p <0.001), as indicated by the Alamar blue assay. The Live/Dead assay also revealed a greater density of viable cells in the laser group. The cell cycle analysis revealed a significant increase in the number of cells in the proliferative phase (G2/M) in the laser group compared with the control group (p <0.001). The SEM images demonstrated that the irradiated group had a greater concentration of cells while still maintaining their cell shape and projections. This study demonstrated that photobiomodulation can increase the viability and proliferation of periodontal stem cells cultured on PLA scaffolds, suggesting the potential of this protocol for future studies on periodontal tissue engineering., (© 2024. The Author(s), under exclusive licence to Springer-Verlag London Ltd., part of Springer Nature.)

Aims: The study aimed to investigate the role of miR-146a-5p in osteogenesis of hPDLSCs irradiated with low-energy red LEDs., Methods: After irradiation with 5 J/cm 2 red LED, miR-146a-5p expression was detected by real-time quantitative polymerase chain reaction (RT-qPCR), and osteogenic markers expression was determined by RT-qPCR and Western blotting. Alkaline phosphatase (ALP) activity was assessed by ALP staining, and mineralization was assessed by Alizarin Red staining, respectively. Lentiviral vectors were designed to regulate miR-146a-5p expression. Dual-luciferase reporter assay was performed to confirm the targeted relationship between miR-146a-5p and MAPK1. Short hairpin RNA (shRNA) was used to regulate MAPK1 expression., Results: RT-qPCR and western blotting revealed that 5 J/cm 2 irradiation elevated the levels of the osteogenic markers osterix (OSX) and bone sialoprotein (BSP) in hPDLSCs. miR-146a-5p is downregulated in hPDLSCs under the low-energy red LED light irradiation. miR-146a-5p underexpression markedly promoted the osteogenic potential of hPDLSCs. miR-146a-5p targeted MAPK1. 5 J/cm 2 red LED irradiation rescued the inhibitory effects of upregulated miR-146a-5p on osteogenic differentiation, and the positive influence of red LED irradiation could be reversed by downregulated MAPK1., Conclusion: These findings confirm that miR-146a-5p is involved in the effect of LED irradiation on the osteogenic differentiation of hPDLSCs by targeting MAPK1. Red LED irradiation may be a potential clinical adjunct therapy for periodontal regeneration., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Shortening retention time and minimizing relapse rates are ongoing challenges in orthodontics. This study investigated the effects of natural fulvic acids (FAs) and low-level laser therapy (LLLT) on orthodontic retention in rats., Methods: Seventy-two male Sprague-Dawley rats underwent mesial movement of the left maxillary first molar using a 50 g force via a nickel-titanium tension spring. After three weeks of movement, the rats entered the retention phase with retainer wires and were divided into four groups: Control (no intervention), FAs (80 mg/kg orally daily), LLLT (808 nm laser twice weekly), and FAs + LLLT (both treatments). Retainers were removed on days 7, 14, and 21 for a 3-day relapse assessment. Maxillary impressions were analyzed for relapse rates using 3Shape software, alongside histological and immunohistochemical evaluations of bone morphogenetic protein-2 (BMP-2) expression in periodontal tissues, with differences among groups analyzed using an ordinary two-way analysis of variance (ANOVA)., Results: The relapse rate decreased over time, particularly at 10, 17, and 24 days (p < 0.001). The FAs group did not significantly affect relapse rates compared to the control group (p = 0.084). In contrast, both the LLLT and FAs + LLLT groups significantly reduced relapse rate (p < 0.001), with no significant difference between these groups (p = 0.555). Histological examination revealed active osteoclasts on day 10, decreasing by days 17 and 24. The LLLT and FAs + LLLT groups showed less local cementum resorption and better periodontal fiber arrangement. All treatment groups significantly increased BMP-2 expression (P < 0.05) compared to controls. with LLLT and FAs + LLLT differing significantly from FAs (P < 0.001), though no difference was observed between LLLT and FAs + LLLT (P = 0.578)., Conclusions: FAs did not significantly reduce relapse rate with retainers, while LLLT effectively reduced relapse rates, showing no additional benefit from combining FAs with LLLT. Both FAs and LLLT increased BMP-2 expression in PDL fibroblasts but with no synergistic effect., (© 2024. The Author(s).)

The present study aimed to compare the effect of photobiomodulation with different energy densities on the angiogenic differentiation of human periodontal ligament stem cells (hPDLSCs) and stem cells from human exfoliated deciduous teeth (SHED). Photobiomodulation therapy with a 660 nm diode laser (2.4 J/cm 2 and 3.9 J/cm 2 ) on two consecutive days post-culture was applied to two types of stem cells (hPDLSCs and SHED). The Quantitative Real-time Polymerase Chain Reaction (RT-qPCR) test was undertaken to investigate Vascular Endothelial Growth Factor-A (VEGF-A) and Angiopoietin I (ANG-I) genes on days 1, 3, 5, 7, and 10 after the first session of laser application. The 4',6-diamidino-2-phenylindole (DAPI) staining and Methyl Thiazolyl Tetrazolium (MTT) test were conducted on days 1, 3, and 5 after the first session of laser application, to assess the cell viability. The Two-way ANOVA with Tukey post hoc test was used to analyze the outcomes of the MTT and RT-qPCR tests. The results of the MTT and DAPI convergently illustrated that the groups receiving photobiomodulation with 2.4 J/cm 2 had higher cell viability compared to 3.9 J/cm 2 . All experimental groups showed an upregulation of VEGF-A and ANG-I gene expression from day 1 to 5, followed by a downregulation from day 5 to 10. The groups with cultured hPDLSCs and SHED receiving photobiomodulation using 2.4 J/cm 2 had the most amounts of VEGF-A and ANG-I gene expression on day 5, respectively. In conclusion, the 660 nm mediated photobiomodulation therapy of cultured SHED and hPDLSCs with 2.4 J/cm 2 energy density may be associated with higher angiogenic differentiation (the expression of VEGF-A and ANG-I) as well as higher cell viability compared to the photobiomodulation therapy with 3.9 J/cm 2 ., (© 2024. The Author(s).)

This study aimed to compare the effects of photobiomodulation therapy (PBMT) with 660 and 980 nm diode lasers on differentiation of periodontal ligament mesenchymal stem cells (PDLMSCs). In this in vitro, experimental study, PDLMSCs were obtained from the Iranian Genetic Bank and cultured in osteogenic medium. They were then subjected to irradiation of 660 and 980 nm diode lasers, and their viability was assessed after one, two, and three irradiation cycles using the methyl thiazolyl tetrazolium (MTT) assay. The cells also underwent DAPI staining, cell apoptosis assay by using the Annexin V/PI, Alizarin Red staining, and real-time polymerase chain reaction (PCR) for assessment of the expression of osteogenic genes. Data were analyzed by two-way ANOVA. The two laser groups had no significant difference in cell apoptosis according to the results of DAPI staining. Both laser groups showed higher cell viability in the MTT assay at 4 and 6 days compared with the control group. Annexin V/PI results showed higher cell viability in both laser groups at 4 days compared with the control group. Rate of early and late apoptosis was lower in both laser groups than the control group at 4 days. Necrosis had a lower frequency in 980 nm laser group than the control group on day 6. Alizarin Red staining showed higher cell differentiation in both laser groups after 3 irradiation cycles than the control group. The highest expression of osteopontin (OPN), osteocalcin (OCN), and Runt-related transcription factor 2 (RUNX2) was noted in 660 nm laser group with 3 irradiation cycles at 14 days, compared with the control group. PBMT with 660 and 980 nm diode lasers decreased apoptosis and significantly increased PDLMSC differentiation after 3 irradiation cycles., (© 2024. The Author(s).)

Periodontal disease is a chronic inflammatory disease caused by periodontal bacteria. Recently, periodontal phototherapy, treatment using various types of lasers, has attracted attention. Photobiomodulation, the biological effect of low-power laser irradiation, has been widely studied. Although many types of lasers are applied in periodontal phototherapy, molecular biological effects of laser irradiation on cells in periodontal tissues are unclear. Here, we have summarized the molecular biological effects of diode, Nd:YAG, Er:YAG, Er,Cr:YSGG, and CO 2 lasers irradiation on cells in periodontal tissues. Photobiomodulation by laser irradiation enhanced cell proliferation and calcification in osteoblasts with altering gene expression. Positive effects were observed in fibroblasts on the proliferation, migration, and secretion of chemokines/cytokines. Laser irradiation suppressed gene expression related to inflammation in osteoblasts, fibroblasts, human periodontal ligament cells (hPDLCs), and endothelial cells. Furthermore, recent studies have revealed that laser irradiation affects cell differentiation in hPDLCs and stem cells. Additionally, some studies have also investigated the effects of laser irradiation on endothelial cells, cementoblasts, epithelial cells, osteoclasts, and osteocytes. The appropriate irradiation power was different for each laser apparatus and targeted cells. Thus, through this review, we tried to shed light on basic research that would ultimately lead to clinical application of periodontal phototherapy in the future.

High-intensity laser therapy (HILT) and photobiomodulation therapy (PBMT) are two types of laser treatment. According to recent clinical reports, PBMT promotes wound healing after trauma or surgery. In addition, basic research has revealed that cell differentiation, proliferation, and activity and subsequent tissue activation and wound healing can be promoted. However, many points remain unclear regarding the mechanisms for wound healing induced by PBMT. Therefore, in this review, we present an example from our study of HILT and PBMT irradiation of tooth extraction wounds using two types of lasers with different characteristics (diode laser and carbon dioxide laser). Then, the effects of PBMT on the wound healing of bone tissues are reviewed from histological, biochemical, and cytological perspectives on the basis of our own study of the extraction socket as well as studies by other researchers. Furthermore, we consider the feasibility of treatment in which PBMT irradiation is applied to stem cells including dental pulp stem cells, the theme of this Special Issue, and we discuss research that has been reported on its effect.

This study aimed to histologically and radiographically evaluate the effectiveness of low-intensity laser irradiation of different wavelengths (660 or 808 nm) as an adjunct to scaling and root planing in the treatment of experimental periodontitis in rats. Periodontitis was induced by placing a ligature around the mandibular first molar of the rats. In total, 40 Wistar rats were randomly divided into five groups (n = 8 each): control (CG), periodontal disease (PD), scaling and root planing (SRP), SRP + 660 nm laser (GL660) and SRP + 808 nm laser (GL808). Groups with laser use received radiation at 6 points in the first molar. The animals were euthanized at baseline and at 7 and 14 days after the interventions. Mandibles were surgically removed for histomorphometric and radiographic assessment of periodontal tissues. The GL660 group showed lesser bone loss than the PD group (P < 0.05) and greater alveolar bone margin after 14 days, indicating a better long-term treatment response (P < 0.05). These findings suggest that SRP with the 660 nm laser as an adjunct results in more favorable radiographic and histological responses than the 808 nm laser.

Objective: To investigate the effects of Er:YAG laser on the attachment of human periodontal ligament fibroblasts (hPDLFs) to denuded root surfaces simulating delayed replantation cases. Background data: Dental avulsion is one of the most severe dental traumas, which is often treated with replantation. In delayed replantation scenarios, poor prognosis, including root resorption, usually occurs due to poor root surface conditioning and nonviable hPDLF attachment. Methods: Thirty-six root fragments (5 × 5 × 2 mm) were obtained from periodontium tissue-free premolar root surfaces. Specimens were randomly and equally assigned to the following: Group A, untreated control; Group B, 25 J/cm 2 and 10 Hz of Er:YAG laser irradiation; and Group C, 50 J/cm 2 and 10 Hz of Er:YAG laser irradiation. Some specimens in each group were then prepared for surface topography visualization under SEM, others were subjected to coculture with hPDLF suspension, and cell adhesion was further evaluated by SEM. Results: Group A presented homogenous smooth root surface, with fewer and round-shaped cells attached; Group B and C exhibited rather rough and irregular morphologies, and spindle-shaped fibroblasts were firmly attached by numerous lamellipodia and extensions. After a 3-day coculture, the number of fibroblasts attached in Group A was significantly lower compared with the other two laser-treated groups ( p  = 0.008 < 0.05). No significant alterations were observed between the two laser groups ( p  = 0.135 > 0.05). Conclusions: Er:YAG laser-treated root surfaces are compatible for the attachment of PDLFs, which suggests that Er:YAG laser irradiation may be used as a promising strategy for root surface conditioning in delayed replantation cases.

Periodontal ligament fibroblasts (PDLFs) have osteogenic capacity, producing bone matrix proteins. Application of bone morphogenic proteins (BMPs) to PDLFs is a promising approach for periodontal regeneration. However, in chronic bone metabolic disorders, such as periodontitis, proper control of accompanying inflammation is essential for optimizing the effects of BMPs on PDLFs. We have previously shown that low-intensity pulsed ultrasound (LIPUS), a medical technology that induces mechanical stress using sound waves, significantly promotes osteogenesis in mesenchymal stem cells. Here, we demonstrate that LIPUS promotes the BMP9-induced osteogenic differentiation of PDLFs. In contrast, BMP2-induced osteogenic differentiation was not altered by LIPUS, probably due to the LIPUS-induced secretion of Noggin, a BMP2 antagonist, from PDLFs. To examine if LIPUS affects inflammatory responses of PDLFs to lipopolysaccharide (LPS) derived from Porphyromonas gingivalis (LPS-PG), we also simultaneously treated PDLFs with LIPUS and LPS-PG. Treatment with LIPUS significantly inhibited the phosphorylation of ERKs, TANK-binding kinase 1, and interferon regulatory factor 3 in LPS-PG-stimulated PDLFs, in addition to inhibiting the degradation of IκB. Furthermore, LIPUS treatment reduced messenger RNA (mRNA) expression of interleukin-1alpha (IL-1alpha), IL-1beta, IL-6, IL-8, C-C motif chemokine ligand 2, C-X-C motif chemokine ligand 1 (CXCL1), CXCL10 and receptor activator of nuclear factor kappa-B ligand, and also diminished IL-1ß and tumor necrosis factor a (TNFa)-induced inflammatory reactions. Phosphorylation of Rho-associated kinase 1 (ROCK1) was induced by LIPUS, while ROCK1-specific inhibitor prevented the promotive effects of LIPUS on p38 phosphorylation, mRNA expression of CXCL1 and Noggin, and osteogenesis. The suppressive effects of LIPUS on LPS-PG-stimulated inflammatory reactions were also prevented by ROCK1 inhibition. Moreover, LIPUS treatment blocked inhibitory effects of LPS-PG and IL-1ß on osteogenesis. These results indicate that LIPUS suppresses inflammatory effects of LPS-PG, IL-1ß, and TNFa and also promotes BMP9-induced osteogenesis through ROCK1 in PDLFs., (© 2019 Wiley Periodicals, Inc.)

Periodontal disease is a chronic inflammatory disease that is commonly treated with surgical and nonsurgical techniques. However, both approaches have limitations. Low-level laser therapy (LLLT) has been widely applied in reducing inflammatory reactions, and research indicates that LLLT induces an anti-inflammatory effect that may enhance periodontal disease therapy. The purpose of this study was to investigate the anti-inflammatory effect of LLLT on human periodontal ligament cells (hPDLCs) in an inflammatory environment and aimed to determine the possible mechanism of action. Cells were cultured and treated with or without lipopolysaccharide (LPS) from Porphryromonas gingivalis or Escherichia coli, followed by irradiation with a gallium-aluminum-arsenide (GaAlAs) laser (660 nm) at an energy density of 8 J/cm 2 . Quantitative real-time polymerase chain reactions were used to assess the expression of pro-inflammatory genes, including tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, and IL-8. The dual-luciferase reporter assay was used to examine nuclear factor-κB (NF-κB) transcriptional activity. An enzyme-linked immunosorbent assay was used to monitor the concentration of intracellular cyclic adenosine monophosphate (cAMP). Both LPS treatments significantly induced the mRNA expression of pro-inflammatory cytokines. However, LLLT inhibited the LPS-induced pro-inflammatory cytokine expression and elevated intracellular levels of cAMP. The LLLT inhibitory effect may function by downregulating NF-κB transcriptional activity and by increasing the intracellular levels of cAMP. LLLT might inhibit LPS-induced inflammation in hPDLCs through cAMP/NF-κB regulation. These results should be further studied to improve periodontal therapy.

The aim of the study was to compare the effectiveness of laser singlet phototherapy and traditional photodynamic therapy the treatment of periodontal diseases in an animal model. The experimental model involved 70 male rats Wistar in which periodontitis was modeled and treated: in group I (30 animals) a nanosecond laser device for medical use with a wavelength of 1270 nm was used for 7 sessions in a 400 ns pulse mode, an average radiation power of 2 W, and a radiation density of 200 J/cm 2 , group II (30 animals) received photodynamic therapy with the administration of a photosensitizer, followed by irradiation with a laser wavelength of 660 nm for 7 sessions 2 Wt average radiation power, group III (controls, 10 animals) - traditional drug therapy. Morphological studies were performed on 7, 14 and 21 day after treatment. On day 7th and 14th the study revealed In group I the presence of full blood vessels and diffuse expressed leukocyte infiltration with an admixture of macrophages, in group II - pronounced edema of the tissue and vasoconstriction. On day 21 the picture included in group I regenerated periodontal ligament with dilated full blood vessels on the border with the bone beams of the alveolar bone, in group II a moderately pronounced edema of the periodontal ligament with single dilated vessels, in controls significantly destroyed periodontal ligament substituted with granulation tissue and periodontal ligament. Thus, the treatment of periodontitis with the methods of singlet phototherapy leads to the development of reactive inflammation and significant vascularization of periodontal tissues which contributes to the rapid regeneration and stability of remission.

Objective: To clarify whether low-intensity pulsed ultrasound (LIPUS) exposure has recovery effects on the hypofunctional periodontal ligament (PDL) and interradicular alveolar bone (IRAB)., Materials and Methods: Twelve-week-old male Sprague-Dawley rats were divided into three groups (n = 5 each): a normal occlusion (C) group, an occlusal hypofunction (H) group, and an occlusal hypofunction group subjected to LIPUS (HL) treatment. Hypofunctional occlusion of the maxillary first molar (M1) of the H and HL groups was induced by the bite-raising technique. Only the HL group was irradiated with LIPUS for 5 days. The IRAB and PDL of M1 were examined by microcomputed tomography (micro-CT) analysis. To quantify mRNA expression of cytokines involved in PDL proliferation and development, real-time reverse transcription quantitative PCR (qRT-PCR) was performed for twist family bHLH transcription factor 1 (Twist1), periostin, and connective tissue growth factor (CTGF) in the PDL samples., Results: Micro-CT analysis showed that the PDL volume was decreased in the H group compared with that of the C and HL groups. Both bone volume per tissue volume (BV/TV) of IRAB was decreased in the H group compared with that in the C group. LIPUS exposure restored BV/TV in the IRAB of the HL group. qRT-PCR analysis showed that Twist1, periostin, and CTGF mRNA levels were decreased in the H group and increased in the HL group., Conclusion: LIPUS exposure reduced the atrophic changes of alveolar bone by inducing the upregulation of periostin and CTGF expression to promote PDL healing after induction of occlusal hypofunction.

Introduction: Mixed outcomes have been found in animal and clinical studies with regard to the use of low-level laser therapy (LLLT) as a modality to accelerate orthodontic tooth movement (OTM). One major reason for the variable findings is the different methodologies and protocols for laser therapy use., Objective: The aim of this study was to determine whether orthodontically moved molars exposed to two different wattages at the same energy density of LLLT exhibited differences in the amount of tooth movement and molecular and histological changes in the adjacent periodontal areas., Methods: An orthodontic force was applied to rat upper first molars exposed to 500mW (EX-500) and 1000mW (EX-1000) of laser application, with a control group (CT) with no laser application. Gene expression in the periodontal ligament (PDL) and histology of the palatal gingiva of the molars were analyzed., Results: There was a statistically significant difference for OTM between EX-500 but not between EX-1000 and CT groups. RANKL and MMP-13 expression levels in the PDL of orthodontically moved molars, however, were increased significantly in laser-exposed groups compared to CT. Early signs of dysplasia were observed in over half of the animals in the EX-1000 group., Conclusions: Our results provide evidence for molecular changes and the potential dysplastic effects of laser on the surrounding soft tissues. Further studies are needed to better identify an optimum laser protocol to maximize the desired effect., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Introduction: Piezo1 and Piezo2 are mechanosensitive membrane ion channels. We hypothesized that Piezo proteins may play a role in transducing ultrasound-associated mechanical signals and activate downstream mitogen-activated protein kinase (MAPK) signaling processes in dental cells. In this study, the expression and role of Piezo channels were investigated in dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PDLSCs) after treatment with low-intensity pulsed ultrasound (LIPUS)., Methods: Cell proliferation was evaluated by bromodeoxyuridine incorporation. Western blots were used to analyze the proliferating cell nuclear antigen as well as the transcription factors c-fos and c-jun. Enzyme-linked immunosorbent assay and Western blotting were used to determine the activation of MAPK after LIPUS treatment. Ruthenium red (RR), a Piezo ion channel blocker, was applied to determine the functional role of Piezo proteins in LIPUS-stimulated cell proliferation and MAPK signaling., Results: Western blotting showed the presence of Piezo1 and Piezo2 in both dental cell types. LIPUS treatment significantly increased the level of the Piezo proteins in DPSCs after 24 hours; however, no significant effects were observed in PDLSCs. Treatment with RR significantly inhibited LIPUS-stimulated DPSC proliferation but not PDLSC proliferation. Extracellular signal-related kinase (ERK) 1/2 MAPK was consistently activated in DPSCs over a 24-hour time period after LIPUS exposure, whereas phosphorylated c-Jun N-terminal kinase and p38 mitogen-activated protein kinase MAPK were mainly increased in PDLSCs. RR affected MAPK signaling in both dental cell types with its most prominent effects on ERK1/2/MAPK phosphorylation levels; the significant inhibition of LIPUS-induced stimulation of ERK1/2 activation in DPSCs by RR suggests that stimulation of DPSC proliferation by LIPUS involves Piezo-mediated regulation of ERK1/2 MAPK signaling., Conclusions: This study for the first time supports the role of Piezo ion channels in transducing the LIPUS response in dental stem cells., (Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

To evaluate one-session endodontic treatment with aPDT and two-session treatment with calcium hydroxide (CH)-based dressing in dog's teeth with apical periodontitis. After experimental induction of apical periodontitis, 48 teeth were randomly assigned to the following groups: groups OS/aPDT120d and OS/aPDT180d (one-session treatment with aPDT) and groups TS/CH120d and TS/CH180d (two-session treatment with CH-based dressing-control groups). The animals were euthanized after 120 and 180 days. After histotechnical processing, microscopic and radiographic analyses were performed. Data were analyzed by Kruskal-Wallis and Fisher's exact tests (α = 0.05). Groups TS/CHs presented repaired resorbed cemental areas, with collagen bundles and few inflammatory cells. In groups OS/aPDTs, the areas of cemental resorption were not repaired with reduced presence of cells and fibers. In the analysis of the apical closure, fluorescence microscopy and percentage of radiographic reduction of lesions, there was significant difference between groups TS/CH120d and OS/aPDT120d and between TS/CH180d and OS/aPDT180d (p < 0.05). Groups TS/CHs had weak RANKL expression and positive immunostaining for RANK and OPG. In OS/aPDT120d, there was positive immunostaining for RANKL. In OS/aPDT180d, the three osteoclastogenesis markers were expressed. The results using aPDT were worse than those obtained with two-session endodontic treatment using a CH-based dressing in teeth with apical periodontitis.

Objectives: Changes to the radiographic appearance of the jaws after head and neck radiotherapy have not been thoroughly characterized. This retrospective study examines changes to the appearance of the mandible on panoramic images following intensity-modulated radiotherapy (IMRT) and relates these changes to medical co-morbidities and radiation dose., Study Design: The medical and dental charts, and panoramic images of 126 patients who received IMRT at the Princess Margaret Hospital between January 1, 2005, and December 31, 2008, were analyzed independently by three observers., Results: Of the 126 patients, 75 (60%) had post-IMRT changes, as seen on panoramic images; most, 66 (88%), consisted of widened periodontal ligament space (WPLS). The median time to WPLS was 29 months after IMRT. Female gender and radiation dose correlated with decreased time to WPLS., Conclusions: These results indicate that WPLS is a common radiographic sequela after head and neck radiotherapy, underscoring its clinical significance as a reliable marker of irradiated bone. Furthermore, this type of WPLS needs to be differentiated from odontogenic inflammatory disease and cancer recurrence to avoid unnecessary treatment that may precipitate osteoradionecrosis., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Introduction: Mesenchymal stem cells (MSCs) from dental tissues may respond to low-intensity pulsed ultrasound (LIPUS) treatment, potentially providing a therapeutic approach to promoting dental tissue regeneration. This work aimed to compare LIPUS effects on the proliferation and MAPK signaling in MSCs from rodent dental pulp stem cells (DPSCs) compared with MSCs from periodontal ligament stem cells (PDLSCs) and bone marrow stem cells (BMSCs)., Methods: Isolated MSCs were treated with 1-MHz LIPUS at an intensity of 250 or 750 mW/cm2 for 5 or 20 minutes. Cell proliferation was evaluated by 5-bromo-2-deoxyuridine (BrdU) staining after 24 hours of culture following a single LIPUS treatment. Specific ELISAs were used to determine the total and activated p38, ERK1/2, and JNK MAPK signaling proteins up to 4 hours after treatment. Selective MAPK inhibitors PD98059 (ERK1/2), SB203580 (p38), and SP600125 (JNK) were used to determine the role of activation of the particular MAPK pathways., Results: The proliferation of all MSC types was significantly increased after LIPUS treatment. LIPUS at a 750-mW/cm2 dose induced the greatest effects on DPSCs. BMSC proliferation was stimulated in equal measures by both intensities, whereas 250 mW/cm2 LIPUS exposure exerted maximum effects on PDLSCs. ERK1/2 was activated immediately in DPSCs after treatment. Concomitantly, DPSC proliferation was specifically modulated by ERK1/2 inhibition, whereas p38 and JNK inhibition exerted no effects. In BMSCs, JNK MAPK signaling was LIPUS activated, and the increase in proliferation was blocked by specific inhibition of the JNK pathway. In PDLSCs, JNK MAPK signaling was activated immediately after LIPUS, whereas p-p38 MAPK increased significantly in these cells 4 hours after exposure. Correspondingly, JNK and p38 inhibition modulated LIPUS-stimulated PDLSC proliferation., Conclusions: LIPUS promoted MSC proliferation in an intensity and cell-specific dependent manner via activation of distinct MAPK pathways., (Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Objective: To investigate combined effect of photobiomodulation with a matrix metalloproteinase (MMP) inhibitor on the relapse rate in relation to MMP expression in rats., Materials and Methods: Fifty-two rats were divided into four groups according to the treatment modality: control group, irradiation group, doxycycline group, and irradiation with doxycycline group. During a relapse period of 5 days after orthodontic movement, maxillary central incisors were treated by low-level laser therapy (LLLT) as a photobiomodulation and/or doxycycline as a synthetic MMP inhibitor. Relapse rate was evaluated in association with MMP expression at the gene and protein levels., Results: Relapse rates were increased by LLLT (1.57-fold) and decreased by doxycycline (0.83-fold) compared with the control, showing positive correlation with the levels of expression for all MMPs in the periodontal ligament (PDL). LLLT concomitant with doxycycline administration resulted in no significant differences of relapse rate and MMP expression from the control., Conclusions: The combined effect of photobiomodulation with an MMP inhibitor around the relapsing teeth proved to be antagonistic to PDL remodeling activity during relapse. This study suggests a basis for developing a novel biologic procedure targeting the MMP-dependent PDL remodeling to control the relapse rate.

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., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Objective: To investigate the mechanisms that cause damage to root formation as a result of irradiation to the mouse head, morphological changes in molar dental roots and cell dynamics in Hertwig's epithelial root sheath (HERS), and surrounding mesenchymal tissue were examined., Materials and Methods: To perform the experiments, 5-day-old C57BL/6 mice were randomly divided into three groups: the control group (0 Gy) and irradiated groups (10 and 20 Gy). Micro-CT analysis, HE staining, immunohistochemistry analysis, and TUNEL assay were then performed., Results: Roots in irradiated mice were dose-dependently shorter than those of control mice. Cells located outside the root dentin, with abnormal morphology in irradiated mice, were positive for an odontoblast marker. HERS fragmentation occurred earlier in irradiated mice than in control mice, and HERS was trapped by the calcified apical tissue. A dose-dependent reduction in the number of proliferating cells within the apical dental pulp and periapical periodontal ligament surrounding HERS was observed in irradiated mice. Apoptotic cells in the dental pulp and periodontal ligament surrounding HERS were hardly seen., Conclusions: These results indicate that the early disappearance of HERS and the proliferative suppression of the surrounding mesenchymal cells, which was induced by irradiation, caused dental root malformation., (© 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Low-level laser has been indicated to have the capability to facilitate the differentiation of the osteoclastic and osteoblastic cells which are responsible for the bone remodeling process. The aim of this study was to evaluate the effects of InGaAlP laser with a wavelength of 660 nm on the rate of tooth movement and histological status. Thirty male Wistar rats of 7 weeks old were selected for this study. The rats were randomly divided into two groups of 15 each to form the experimental (laser-irradiated) and control (non-irradiated) groups. The control group received unilateral orthodontic appliance design (one quadrant), but the laser-irradiated group received split-mouth design, with orthodontic appliance on both sides and laser irradiation on one side only (group b) and on the contralateral side (group c). The orthodontic appliance consisted of a NiTi closed coil spring with a length of 5 mm which was ligated to maxillary molar and incisor. A total of 60 g of force was applied to the rat molar. The diode laser (660 nm) was irradiated with an output power of 25 mW in continuous mode for a total time of 5 min in the laser-irradiated group. After 14 days of orthodontic tooth movement, the amount of tooth movements was measured. In the laser-irradiated group, the amount of tooth movement was significantly greater than that of the non-irradiated group (2.3-fold), but there was no significant difference between the non-irradiated and indirectly irradiated groups. Histopathological studies revealed that the number of osteoclasts in the laser-irradiated group was significantly greater than that of the non-irradiated group (1.5-fold) while this number was almost the same in the non-irradiated and indirectly irradiated groups. The results suggested that low-level laser can accelerate the rate of bone remodeling. However, in order to utilize the low-level laser as an adjunct in orthodontic practice on patients, further research studies are needed for finding the appropriate dosage for the human tissues.

Introduction: In this study, we compared the effectiveness of laser-aided circumferential supracrestal fiberotomy (CSF) and low-level laser therapy (LLLT) with conventional CSF in reducing relapse of corrected rotations., Methods: The study included 24 patients who were at the finishing stage of orthodontic treatment and had at least 1 maxillary incisor with 30° to 70° of rotation before starting therapy. The subjects were divided into 4 groups by treatment: conventional CSF, Er:YAG laser-aided CSF, LLLT, and control. After alginate impressions were taken, the archwire was sectioned from the experimental incisors, and they were allowed to relapse. The second impression was taken 1 month later, and the degree and percentage of relapse were calculated in photographs taken from the dental models. Gingival recession, pocket depth, and pain were also measured in the CSF groups., Results: The mean percentages of relapse were 9.7% in the conventional CSF, 12.7% in the Er:YAG laser-aided CSF, 11.7% in the LLLT, and 27.8% in the control groups. Relapse was significantly greater in the control than the experimental groups (P <0.05), which were not statistically different from each other. The changes in sulcus depth and gingival recession were small and not significantly different among the CSF groups (P >0.05), but pain intensity was greater in subjects who underwent conventional CSF (P = 0.003)., Conclusions: Er:YAG laser-aided CSF proved to be an effective alternative to conventional CSF in reducing rotational relapse. LLLT with excessively high energy density was also as effective as the CSF procedures in alleviating relapse, at least in the short term., (Copyright © 2014 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Malocclusions, such as an open bite and high canines, are often encountered in orthodontic practice. Teeth without occlusal stimuli are known as hypofunctional teeth, and numerous atrophic changes have been reported in the periodontal tissue, including reductions in blood vessels in the periodontal ligament (PDL), heavy root resorption, and reduced bone mineral density (BMD) in the alveolar bone. Low Level Laser (LLL) has been shown to have a positive effect on bone formation and the vasculature. Although the recovery of hypofunctional teeth remains unclear, LLL is expected to have a positive influence on periodontal tissue in occlusal hypofunction. The aim of the present study was to elucidate the relationship between LLL and periodontal tissue in occlusal hypofunction. Twenty-four male rats aged 5 weeks were randomly divided into control and hypofunctional groups. An anterior metal cap and bite plate were attached to the maxillary and mandibular incisors in the hypofunctional group to simulate occlusal hypofunction in the molars. LLL irradiation was applied to the maxillary first molar through the gingival sulcus in half of the rats. Rats were divided into four groups; control, control+LLL, hypofunctional, and hypofunctional+LLL. Exposure to LLL irradiation was performed for 3 minutes every other day for 2 weeks. Animals were examined by Micro-CT at 5 and 7 weeks and were subsequently sacrificed. Heads were resected and examined histologically and immunohistologically. The hypofunctional group had obvious stricture of the PDL. However, no significant differences were observed in the PDL and alveolar bone between the hypofunctional+LLL and the control groups. In addition, the expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF)-positive cells were higher in the hypofunctional + LLL group than in the hypofunctional group. These results indicated that LLL enhanced the production of bFGF and VEGF in the periodontal tissue of hypofunctional teeth.

Objective: The purpose of this research was to evaluate the histological changes of the periodontal ligament and alveolar bone during dental movement in diabetic rats subjected to low level laser therapy (LLLT)., Methods: The movement of the upper molar was performed in 60 male Wistar rats divided into four groups (n=15): CTR (control), DBT (diabetic), CTR/LT (irradiated control) and DBT/LT (irradiated diabetic). Diabetes was induced with alloxan (150 mg/kg, i.p.). LLLT was applied with GaAlAs laser at 780 nm (35 J/cm(2)). After 7, 13 and 19 days, the periodontal ligament and alveolar bone were histologically analyzed., Results: The mean of osteoblasts (p<0.01) and blood vessels (p<0.05) were significantly decreased in DBT compared with CTR at 7 days, whereas the mean of osteoclasts was lower at 7 (p<0.001) and 13 days (p<0.05). In DBT/LT, only the mean of osteoclasts was lower than in CTR (p<0.05) at 7 days, but no difference was observed at 13 and 19 days (p>0.05). The collagenization of the periodontal ligament was impaired in DBT, whereas DBT/LLT showed density/disposition of the collagen fibers similar to those observed in CTR., Conclusions: LLLT improved the periodontal ligament and alveolar bone remodeling activity in diabetic rats during dental movement., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objectives: Low-intensity pulsed ultrasound (US) can accelerate fracture healing and osteogenic differentiation. The aim of this study was to investigate the osteogenic effect of low-intensity pulsed US on human periodontal ligament cells and to determine whether bone morphogenetic protein (BMP)-Smad signaling was involved., Methods: Human periodontal ligament cells were exposed to low-intensity pulsed US at a frequency of 1.5 MHz and intensity of 90 mW/cm(2) for 20 min/d. Osteogenic differentiation was determined by assaying alkaline phosphatase (ALP) and calcium deposition. Expression of BMP-2, BMP-6, and BMP-9 was detected by real-time polymerase chain reaction analysis. Phosphorylated Smad was detected by western blotting; Smad in the cells was labeled by an immunofluorescent antibody and observed by laser-scanning confocal microscopy., Results: The optical density of ALP stimulated by US at 1.5 MHz and 90 mW/cm(2) for 20 min/d was significantly higher than in other groups (P < .01); therefore, this dosage was considered optimal for promoting osteogenic differentiation. After 13 days of US exposure, ALP increased gradually after 5 days, peaked at 11 days, and decreased at 13 days, with a significant difference compared with the control group (P < .05). Osteocalcin production increased from 9 to 13 days and peaked at 15 days, with a significant difference compared with the control group (P < .05). BMP-2 and BMP-6 increased dynamically after exposure for 13 days. BMP-2 increased 6.07-fold at 3 days, 6.39-fold at 11 days, and 5.97-fold at 13 days. BMP-6 expression increased 6.82-fold at 1 day and 51.5-fold at 3 days and decreased thereafter. BMP-9 was not expressed. Phospho-Smad1/5/8 expression was significantly increased after exposure (P< .05) and transferred from the cytoplasm into the nuclei., Conclusions: Low-intensity pulsed US effectively induced osteogenic differentiation of human periodontal ligament cells, and the BMP-Smad signaling pathway was involved in the mechanism.

Loss of teeth increases with age or after genotoxic treatments, like head and neck radiotherapy, due to periodontium breakdown. Periodontal ligament fibroblasts represent the main cell type in this tissue and are crucial for the maintenance of homeodynamics and for its regeneration. Here, we have studied the characteristics of human periodontal ligament fibroblasts (hPDLF) that became senescent after replicative exhaustion or after exposure to ionizing radiation, as well as their ability for osteoblastic differentiation. We found that senescent hPDLF express classical markers of senescence, as well as a catabolic phenotype, as shown by the decrease in collagen type I and the increase of MMP-2 expression. In addition, we observed a considerably decreased expression of the major transcription factor for osteoblastic differentiation, i.e. Runx2, a down-regulation which was found to be p53-dependent. In accordance to the above, senescent cells have a significantly decreased alkaline phosphatase gene expression and activity, as well as a reduced ability for osteoblastic differentiation, as found by Alizarin Red staining. Interestingly, cells from both type of senescence express similar characteristics, implying analogous functions in vivo. In conclusion, senescent hPDLF express a catabolic phenotype and express a significantly decreased ability towards an osteoblastic differentiation, thus probably affecting tissue development and integrity.

Purpose: This study evaluates early changes in human mesenchymal stem cells (MSC) isolated from dental pulp and periodontal ligament after γ-irradiation and the effect of ataxia-telangiectasia mutated (ATM) inhibition., Methods: MSC were irradiated with 2 and 20 Gy by (60)Co. For ATM inhibition, specific inhibitor KU55933 was used. DNA damage was measured by Comet assay and γH2AX detection. Cell cycle distribution and proteins responding to DNA damage were analyzed 2-72 h after the irradiation., Results: The irradiation of MSC causes an increase in γH2AX; the phosphorylation was ATM-dependent. Irradiation activates ATM kinase, and the level of p53 protein is increased due to its phosphorylation on serine15. While this phosphorylation of p53 is ATM-dependent in MSC, the increase in p53 was not prevented by ATM inhibition. A similar trend was observed for Chk1 and Chk2. The increase in p21 is greater without ATM inhibition. ATM inhibition also does not fully abrogate the accumulation of irradiated MSC in the G2-phase of the cell-cycle., Conclusions: In irradiated MSC, double-strand breaks are tagged quickly by γH2AX in an ATM-dependent manner. Although phosphorylations of p53(ser15), Chk1(ser345) and Chk2(thr68) are ATM-dependent, the overall amount of these proteins increases when ATM is inhibited. In both types of MSC, ATM-independent mechanisms for cell-cycle arrest in the G2-phase are triggered.

Retaining or improving periodontal ligament (PDL) function is crucial for restoring periodontal defects. The aim of this study was to evaluate the physiological effects of low-power laser irradiation (LPLI) on the proliferation and osteogenic differentiation of human PDL (hPDL) cells. Cultured hPDL cells were irradiated (660 nm) daily with doses of 0, 1, 2 or 4 J⋅cm(-2). Cell proliferation was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, and the effect of LPLI on osteogenic differentiation was assessed by Alizarin Red S staining and alkaline phosphatase (ALP) activity. Additionally, osteogenic marker gene expression was confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR). Our data showed that LPLI at a dose of 2 J⋅cm(-2) significantly promoted hPDL cell proliferation at days 3 and 5. In addition, LPLI at energy doses of 2 and 4 J⋅cm(-2) showed potential osteogenic capacity, as it stimulated ALP activity, calcium deposition, and osteogenic gene expression. We also showed that cyclic adenosine monophosphate (cAMP) is a critical regulator of the LPLI-mediated effects on hPDL cells. This study shows that LPLI can promote the proliferation and osteogenic differentiation of hPDL cells. These results suggest the potential use of LPLI in clinical applications for periodontal tissue regeneration.

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., (© 2012 John Wiley & Sons A/S.)

Introduction: Different low-level laser (LLL) irradiation protocols have been tested to accelerate orthodontic tooth movement (OTM). Nevertheless, divergent results have been obtained. It was suggested that the stimulatory action of low level laser irradiation occurs during the proliferation and differentiation stages of bone cellular precursors, but not during later stages., Objective: The purpose of this study was to determine the effect of two protocols of LLL irradiation on experimental tooth movement: One with daily irradiations and another with irradiations during the early stages., Methods: Thirty-six rats were divided into control groups (CG1, CG2, CG3) and irradiated groups (IrG1, IrG2, IrG3) according to the presence of: experimental tooth movement, laser irradiation, type of laser irradiation protocol and date of euthanasia (3th or 8th day of experiment). At the end of experimental periods, a quantitative evaluation of the amount of OTM was made and the reactions of the periodontium were analyzed by describing cellular and tissue reactions and by counting blood vessels., Results: The amount of OTM revealed no significant differences between groups in the same experimental period (p < 0.05). Qualitative analysis revealed the strongest resorption activity in irradiated groups after seven days, especially when using the daily irradiation protocol. There was a higher number of blood vessels in irradiated animals than in animals without orthodontic devices and without laser irradiation (p < 0.05)., Conclusion: Moreover, angiogenesis was verified in some of the irradiated groups. The irradiation protocols tested were not able to accelerate OTM and root resorption was observed while they were applied.

This study was designed to investigate the effects of low-intensity laser therapy (LILT) on periodontal ligament (PDL) remodeling during relapse and retention after the completion of orthodontic movement. The maxillary central incisors (n = 104) of the 52 rats were randomly divided into five groups according to the treatment modality: baseline control group without any intervention (n = 8); relapse group without retainer after tooth movement (n = 24); retention group with fixed retainer after tooth movement (n = 24); lased relapse group without retainer after tooth movement and LILT (n = 24); lased retention group with retainer after tooth movement and LILT (n = 24). LILT was daily performed using a gallium-aluminum-arsenide diode laser in a biostimulation mode: wavelength of 780 nm, continuous waves at 70 mW output power, a preset low intensity of 1.75 W/cm(2) in contact mode, resulting in energy dose of 5 J/cm(2) per irradiation for 3 s. The animals were euthanized on days 1, 3, and 7 after removal of the orthodontic appliance. Real-time RT-PCR was performed for quantitative analysis of matrix metalloproteinases mRNA expression. Immunoreactivities of collagen and tissue inhibitor of metalloproteinase were observed on the compression and tension sides. LILT significantly facilitated the expression of five tested MMP mRNAs in both relapse and retention groups. TIMP-1 immunoreactivity was inhibited by LILT in both groups, whereas Col-I immunoreactivity was increased by LILT only in the retention group. These results indicate that LILT would act differently on the stability after orthodontic treatment according to additional retainer wearing or not. LILT when combined with a retainer on the moved teeth may shorten the retention period by accelerating periodontal remodeling in the new tooth position, whereas, LILT on the moved teeth left without any retainer would rather increase the rate of relapse after treatment.

Low-level laser therapy is a tool employed in the management of post-operative inflammation process and in the enhancement of reparative process. The aim of the study was to perform histological evaluation of dental and periodontal ligament of rats central upper-left incisor teeth re-implanted and irradiated with low-level laser (InGaAl, 685 nm, 50 J/cm(2)) 15, 30, and 60 days after re-implantation. Seventy-two male rats had the central upper left incisor removed and kept for 15 min on dry gauze before replantation. Laser was irradiated over the root surface and empty alveolus prior replantation and over surrounding mucosa after the re-implantation. After histological procedures, all slices were analyzed regarding external resorption area and histological aspects. We observed an increase of root resorption (p < 0.05) in the control group compared to the laser group at 15, 30, and 60 days. These results showed that the laser groups developed less root resorption areas than the control group in all experimental periods. Additionally, histological analysis revealed less inflammatory cells and necrotic areas in laser groups.

Objective: We aimed to assess histologic changes after the use of laser phototherapy (LPT) during induced tooth movement with 40 g/F on young adult male rats., Background Data: Hyalinization is a sterile necrosis at the pressure zone of the periodontal ligament observed during the initial stages of the orthodontic movement, and extensive hyaline areas might cause an important delay in the tooth movement. The use of LPT is considered an enhancement factor for bone repair, as it stimulates microcirculation as well as the cellular metabolism., Materials and Methods: Thirty animals were divided into two groups (n=15), named according to the time of animal death (7, 13, and 19 days). Half of the animals in each group were subjected to irradiation with infrared (IR) laser (λ790 nm, round shaped beam, 40 mW, continuous wave (CW), diameter=2 mm (0.0314 cm(2)), 1.273 W/cm(2), time=2×112 sec+1×275 sec (total time 499 sec), 2×142.6/4.48 J+1×350/11 J, 635.2 J/cm(2)/20 J/ session), during orthodontic movement, the other half were used as nonirradiated controls. After animal death, specimens were sectioned, processed, and stained with hematoxylin and eosin (HE) and Sirius Red, and were used for semi-quantitative histologic analysis by light microscopy. Data were statistically analyzed., Results: We demonstrated that LPT positively affected an important aspect of dental movement; the hyalinization. In the present study, we found a significant reduced expression of hyalinization after 19 days. On irradiated subjects, hyalinization was increased at day 7 with significant reduction at day 13., Conclusions: It is possible to conclude that the use of laser light caused histologic alterations during the orthodontic movement characterized by increased formation of areas of hyalinization at early stages, and late reduction when compared to nonirradiated animals.

Background: We have recently reported that a low level of CO₂ laser irradiation induced growth stimulation (hormesis) of both human gingival fibroblast (HGF) and oral squamous cell carcinoma cell line (HSC-2), but the extent of hormetic response was much smaller than that previously reported for toxicants and radiation in other experimental systems. Here we investigated the extent of hormetic response induced by CO₂ laser irradiation in human pulp cells (HPCs) and periodontal ligament fibroblast (HPLF)., Materials and Methods: HPC and HPLF cells were established from the periodontal tissues of the first premolar extracted tooth. Cells were cultured for 24, 48 or 72 hours after exposure to various irradiation powers, and the viable cell number was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method., Results: CO₂ laser irradiation induced biphasic effects on the growth of both HPC and HPLF cells. The maximum hormetic response was less than 50%. The hormetic response was found within the energy density of 7.98-79.77 J/cm², and cytotoxicity emerged at powers over 132.96 J/cm². Combining with our previous report, HPCs showed the highest hormetic response, followed by HPLFs and then HGFs. Both HPLFs and HGFs showed similar time-course of hormesis response, increasing response with incubation time., Conclusion: The hormetic response may be the common survival mechanism by which cells escape from radiation-induced injury. Higher hormetic response of HPCs may reflect their potential for differentiation into one of the components in dentin.

The purpose of this study was to establish a long-term tooth cryopreservation method that can be used for tooth autotransplantation. Human periodontal ligament (PDL) cells were frozen in 10% dimethyl sulfoxide (Me(2)SO) using a programmed freezer with a magnetic field. Cells were cryopreserved for 7 days at -150 degrees C. Immediately after thawing, the number of surviving cells was counted and the cells were cultured; cultured cells were examined after 48 h. Results indicated that a 0.01 mT of a magnetic field, a 15-min hold-time, and a plunging temperature of -30 degrees C led to the greatest survival rate of PDL cells. Based on these findings, whole teeth were cryopreserved under the same conditions for 1 year. The organ culture revealed that the PDL cells of cryopreserved tooth with a magnetic field could proliferate as much as a fresh tooth, although the cells did not appear in the cryopreserved tooth without a magnetic field. Histological examination and the transmission electron microscopic image of cryopreserved tooth with a magnetic field did not show any destruction of cryopreserved cells. In contrast, severe cell damage was seen in cells frozen without a magnetic field. These results indicated that a magnetic field programmed freezer is available for tooth cryopreservation., ((c) 2010 Elsevier Inc. All rights reserved.)

Background and Objective: The therapeutic rationale of low-energy pulsed CO(2) laser coagulation mode has not been clarified yet. We conducted this study to characterize the effect of low-energy pulsed CO(2) laser coagulation mode irradiation of the rat gingiva in terms of the expression of heat shock proteins., Material and Methods: Laser irradiation was achieved with the parameters of 5 W, 600 mus pulse duration, and fluence of 326 J/cm(2). The gingiva dissected at different times after irradiation was processed for immunohistochemical examination of the expression of the heat shock proteins, Hsp70 and Hsp25., Results: One hour after irradiation, the epithelial keratinocytes facing the laser wound exhibited an overexpression of Hsp70 in their nucleus. The connective tissue cells facing the laser wound, which included fibroblasts and capillary endothelial cells, showed de novo expression of Hsp70 at 3 h post-irradiation, the level of which peaked at 1 d and thereafter decreased. An enhanced and/or de novo expression of Hsp25 in the connective tissue cells facing the laser wound became evident at 3 h after irradiation, and after 1 d the Hsp25-expressing cells increased in number and spread over the wound as wound repair progressed. There was a temporospatial difference in the expression pattern between Hsp70 and Hsp25, with only a few cells appearing to co-express both heat shock proteins., Conclusion: The CO(2) laser treatment in coagulation mode produced the expression of heat shock proteins, and the findings suggest that while Hsp70 mainly conferred cell protection, Hsp25 was involved in the progress of wound repair as well as cell protection.

Patients often feel pain or discomfort in response to orthodontic force. It was hypothesized that CO(2) laser irradiation may reduce the early responses to nociceptive stimuli during tooth movement. The distribution of Fos-immunoreactive (Fos-IR) neurons in the medullary dorsal horn of rats was evaluated. Two hrs after tooth movement, Fos-IR neurons in the ipsilateral part of the medullary dorsal horn increased significantly. CO(2) laser irradiation to the gingiva just after tooth movement caused a significant decrease of Fos-IR neurons. PGP 9.5- and CGRP-positive nerve fibers were observed in the PDL of all study groups. The maximum temperature below the mucosa during CO(2) laser irradiation was less than 40 degrees C. It was suggested that CO(2) laser irradiation reduced the early responses to nociceptive stimuli during tooth movement and might not have adverse effects on periodontal tissue.

Background: The erbium-doped:yttrium, aluminum, and garnet (Er:YAG) laser has been shown to be a promising tool for root treatment in periodontitis, but little information is available regarding the surface characteristics after this treatment, mainly because it is difficult to obtain standardized dentin samples for in vitro studies., Methods: Commercially available standardized dentin disks were treated with an Er:YAG laser at different settings and used as a substrate for human primary osteoblastic cells (hOBs) and periodontal ligament fibroblasts (PLFs). Cell proliferation on untreated dentin was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after 3, 6, 12, 24, and 48 hours of culture. The effects of the laser on dentin and cell morphology on treated and untreated samples were investigated by scanning electron microscopy after 3, 6, 24, and 48 hours of culture., Results: Dentin samples supported proliferation for both cell types, although growth kinetics were different. The laser dramatically affected the dentin profile, creating a rough and irregular surface. Cells grew easily on untreated dentin, but fewer cells were present on treated areas, often displaying long filopodes. hOBs showed poorer adhesion to treated dentin than PLFs., Conclusions: The dentin disks provide a standardized and useful tool to study dentin surface modifications in vitro. PLFs behaved differently from hOBs on dentin, possibly because of their different affinity to this tissue and/or their differentiation state. The changes induced by the laser produced a less favorable environment for cell adhesion or growth, and treated dentin seemed to be more suitable for PLF adhesion compared to hOB adhesion.

Magnetic attachments have flux leakages, thus they will exert certain magnetic fields on the adjacent tissues when used in the patients' oral cavities. There are few research reports on the biological effects of the magnetic fields generated by magnetic attachments on human body. A cellular static magnetic field (SMF) loading system was developed in this study. By using this system, in vitro cultured human periodontal ligament cells (HPDLCs) were loaded with SMF simulating those of magnetic attachments. The cellular SMF loading system could produce constant SMF and the strength of the SMF is adjustable. The system is small and is able to exert SMF to cells cultured in different culture vessels such as culture dishes and culture plates, thus is suitable to researches in multiple biological items of cells. The results of the SMF loading experiment on HPDLCs showed that this cellular SMF loading system could effectively load cells with SMF of different strengths for different time in vitro. The development of this system has provided a useful tool for the researches on the cellular hiologioal effects of SMF.

Background and Objectives: The purpose of this study was to investigate the long-term effect of Nd:YAG laser irradiation on cultured human periodontal fibroblasts (hPF)., Study Design/materials and Methods: The cultured hPF were irradiated by pulsed Nd:YAG laser. The power delivery was 50 mJ x 10 pps (pulse per second) with irradiation duration 60, 120, 180, or 240 seconds. The viability and collagen content of laser-irradiated hPF were assessed on day 5 after laser treatment. Light microscope and transmission electron microscope (TEM) were used to observe cytomorphological change. The irradiated hPF cultured in mineralizing medium for 28 days were examined by alizarin red S and Von Kossa stain., Results: The cellular viability and collagen content of hPF decreased after Nd:YAG laser irradiation. Cell damage was noted with retraction of cellular processes, loss of normal architecture, and lysis of some cells. However, survived hPF proliferated and migrated to the cell-debris-associated deposits. The electron-dense cytoplasm and amorphous organelles in laser-damaged cells was revealed by TEM. In vitro mineralization was demonstrated in the long-term laser-irradiated hPF cultured in mineralizing medium., Conclusion: Nd:YAG laser irradiation induced partial loss of cellular viability and collagen content. The co-existence of viable cells and progressive degeneration of laser-damaged cells was associated with the in vitro mineralization of hPF., (Copyright 2005 Wiley-Liss, Inc.)

Purpose: The purpose of this study is to evaluate the effect of static magnetic field on the expression of BMP-2 in the periodontal membrane of rat., Methods: 35 female rats of wistar were randomly divided into three groups, which were the normal group, experimental control group and experimental group. The experimental rat's cheeks were put into magnet with the intensity of magnetic field being 0.12 tesla. The rats were sacrificed at 2 days, 4 days, 7 days, respectively. Immunohistochemical assays were used to evaluate the changes of BMP-2 expression in the periodontal membrane of the maxillary molars., Results: BMP-2 was mainly found in the plasma of fibroblast, osteoblast, cementocyte, odontoblast separately. There were more BMP-2 in the experimental group treated with static magnetic field. There were significant differences between the experiment group of 4 day and 7 day and the control group., Conclusions: Static magnetic field stimulated BMP-2 secreted by the cell of periodontal membrane and played an important role in repairing and remodeling of periodontitis.

Objectives: The effect of CO2 laser irradiation on PDL attachment to resected root ends was evaluated., Study Design: Forty dentin wafers were divided into 4 equal groups. Groups I and II were not subjected to laser irradiation. Groups III and IV were subjected to irradiation with CO2 laser (wave length 10.6 mm). PDL cells attachment to the resected root surface (lased/non lased) and cell attachment to the root end filling material (gutta percha and MTA) were evaluated utilizing immunoflouresence microscopy and SEM., Results: PDL cell attachment was observed on dentin surfaces (control samples) and dentin away from the laser strikes (irradiated samples). PDL cell attachment was observed on MTA but was absent on gutta percha., Conclusion: Absence of PDL attachment to laser irradiated dentin could be attributed to morphological surface changes in dentin produced by laser irradiation.

Aim: The aim of this in vitro study was to evaluate a potential stimulatory effect of low-level laser irradiation on the proliferation of human periodontal ligament fibroblasts (PDLF)., Materials and Methods: PDLF obtained from third molar periodontal ligaments were cultured under standard conditions and spread on 96-well tissue culture plates. Subconfluent monolayers were irradiated with an 809-nm diode laser operated at a power output of 10 mW in the continuous wave (cw) mode at energy fluences of 1.96-7.84 Jcm-2. The variable irradiation parameters were the time of exposure (75-300 s per well) and the number of irradiations (1-3). After laser treatment, the cultures were incubated for 24 h. The proliferation rate of the lased and control cultures was determined by means of fluorescence activity of a reduction-oxidation (REDOX) indicator (Alamar Blue Assay) added to the cell culture. Proliferation, expressed in relative fluorescence units (RFU), was determined 24, 48 and 72 h after irradiation., Results: The irradiated cells revealed a considerably higher proliferation activity than the controls. The differences were significant up to 72 h after irradiation (Mann-Whitney U-test, p<0.05)., Conclusion: A cellular effect of the soft laser application is clearly discernible. Clinical studies are needed to evaluate whether the application of low-level laser therapy might be beneficial in regenerative periodontal therapy.

Objective: The purpose of this study was to investigate the effects of low power laser on basic fibroblast growth factors (bFGF) expression in periodontal tissue during tooth movement., Methods: 18 white rabbits were randomly divided into 6 groups with 3 rabbits in each group, including groups of 1, 3, 5, 7, 14 and 21 days. Under an anesthesia condition by 2% pentobarbital sodium, the stainless coil springs were fixed between the first maxillary molar and the incisor producing the force of 80 g. The right side of maxilla was considered as the experimental group under the irradiation of low power laser with the left side as the control groups. The expression of bFGF was investigated half-quantitatively through immunohistochemical analysis., Results: The expression of bFGF in periodontal tissue with irradiation of low power laser was higher than the control side. There were significant differences among the 5, 7, and 14 day groups. In the tension area of the experimental side, the expression of bFGF in the osteoblastic surface of alveolar bone was characteristically greater than that of the control side., Conclusion: The laser of low power promotes the expression of bFGF in the periodontal tissue and alveolar bone remodeling.

Background: The present study is part of a basic research program investigating the cellular effects of an 810 nm GaAlAs-diode laser on human periodontal tissues. The aim of the investigation was to evaluate the effects of laser treatment of root surface specimens on the attachment of periodontal ligament (PDL) cells in vitro., Methods: Root specimens were prepared from periodontally diseased teeth. PDL cells were obtained from human third molar ligaments. Cells were cultured under simple, standardized, and reproducible experimental conditions. One hundred fifty root specimens were scaled and root planed with curets followed by air-powder abrasive treatment; 75 were then lased and 75 served as controls. The irradiation time was 20 seconds at a power output of 1 W. The root segments were placed into culture dishes, covered with a solution of PDL cells, and incubated for 72 hours. The specimens were then washed with phosphate buffer to remove cells not attached to the surface, and the adherent cells were stained with methylene blue. Cells were counted using a reflected light microscope and the cell density per mm2 was calculated., Results: The analysis of 150 specimens revealed no significant differences between the groups (P = 0.347, Wilcoxon test). The cell numbers, however, were slightly higher on laser specimens. The mean was 66 cells/mm2 in the laser group and 63.7 cells/mm2 in the control group., Conclusions: The application of the diode laser at the parameters used did not have a substantially positive effect on the new attachment of PDL cells on the tooth specimens. It remains to be investigated whether the difference detected is really clinically relevant.

Infrared lasers have been used for debridement and sterilisation of both soft and hard tissues, but there have been few studies of such laser applications in endodontics. The present laboratory study was undertaken to examine the feasibility of using pulsed infrared laser radiation to remove moisture from root canals (with an adjunctive sterilising effect). Canals were prepared in extracted teeth and a standardised technique used to fill the apical half of the root canal with saline. Pulses of CO2 or Nd:YAG laser energy were delivered into the root canal system via miniature applicators and residual fluid determined, as well as temperature changes on the root surface. With the CO2 laser, long pulse durations were effective at dehydrating the canals, but elicited deleterious thermal changes both locally within the canal as well as on the root surface. With Nd:YAG laser treatment, large temperature increases on the root surface occurred even with low powers and low pulse frequencies, and extended times were necessary for dehydration. With higher powers and pulse frequencies, complete dehydration could be achieved in less than 60 seconds, however root surface temperatures increased approximately 25 degrees, and the radicular dentine was damaged by the production of plasma. Dehydration of root canals could not be achieved safely with these two infrared lasers, and damage to both radicular dentine and the periodontal ligament would occur if these techniques were to be applied clinically. Alternative methods which do not exert significant thermal effects should be investigated.