In Vitro Cytological Responses against Laser Photobiomodulation for Periodontal Regeneration.

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 CO2 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.

Wound Healing and Cell Dynamics Including Mesenchymal and Dental Pulp Stem Cells Induced by Photobiomodulation Therapy: An Example of Socket-Preserving Effects after Tooth Extraction in Rats and a Literature Review.

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.

Effect of photobiomodulation therapy as an adjunct to scaling and root planing in a rat model of ligature-induced periodontitis: a histological and radiographic study.

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.

Effects of Er:YAG Laser on the Attachment of Human Periodontal Ligament Fibroblasts to Denuded Root Surfaces Simulating Delayed Replantation Cases: An In Vitro Study.

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/cm2 and 10 Hz of Er:YAG laser irradiation; and Group C, 50 J/cm2 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.

Low-intensity pulsed ultrasound promotes bone morphogenic protein 9-induced osteogenesis and suppresses inhibitory effects of inflammatory cytokines on cellular responses via Rho-associated kinase 1 in human periodontal ligament fibroblasts.

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.

[Morphological evaluation of singlet phototherapy in the treatment of periodontal diseases in an experimental study]. / Morfologicheskaia otsenka singletnoi fotooksiterapii pri lechenii zabolevanii parodonta v éksperimental'nom issledovanii.

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/cm2, 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.

Anti-inflammatory effects of low-level laser therapy on human periodontal ligament cells: in vitro study.

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/cm2. 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)-1ß, 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.

Low-intensity pulsed ultrasound reduces periodontal atrophy in occlusal hypofunctional teeth.

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.

Role of Piezo Channels in Ultrasound-stimulated Dental Stem Cells.

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.

Effects of two wattages of low-level laser therapy on orthodontic tooth movement.

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.

Comparison between one-session root canal treatment with aPDT and two-session treatment with calcium hydroxide-based antibacterial dressing, in dog's teeth with apical periodontitis.

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.

Mandibular changes on panoramic imaging after head and neck radiotherapy.

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.

Ultrasound Stimulation of Different Dental Stem Cell Populations: Role of Mitogen-activated Protein Kinase Signaling.

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.

Combined effect of photobiomodulation with a matrix metalloproteinase inhibitor on the rate of relapse in rats.

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.

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.

The effects of diode laser (660 nm) on the rate of tooth movements: an animal study.

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.

Cell dynamics in Hertwig's epithelial root sheath and surrounding mesenchyme in mice irradiated to the head.

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.

Effectiveness of Er:YAG laser-aided fiberotomy and low-level laser therapy in alleviating relapse of rotated incisors.

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.

Effects of a low level laser on periodontal tissue in hypofunctional teeth.

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.

Histological analysis of the periodontal ligament and alveolar bone during dental movement in diabetic rats subjected to low-level laser therapy.

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.