In vitro and clinical evaluation of optical coherence tomography for the detection of subgingival calculus and root cementum.

This study evaluated the effectiveness of swept-source optical coherence tomography (ss-OCT) for detecting calculus and root cementum during periodontal therapy. Optical coherence tomography (OCT) images were taken before and after removal of subgingival calculus from extracted teeth and compared with non-decalcified histological sections. Porcine gingival sheets of various thicknesses were applied to the root surfaces of extracted teeth with calculus and OCT images were taken. OCT images were also taken before and after scaling and root planing (SRP) in human patients. In vitro, calculus was clearly detected as a white-gray amorphous structure on the root surface, which disappeared after removal. Cementum was identified as a thin, dark-gray layer. The calculus could not be clearly observed when soft tissues were present on the root surface. Clinically, supragingival calculus and cementum could be detected clearly with OCT, and subgingival calculus in the buccal cervical area of the anterior and premolar teeth was identified, which disappeared after SRP. Digital processing of the original OCT images was useful for clarifying the calculus. In conclusion, ss-OCT showed potential as a periodontal diagnostic tool for detecting cementum and subgingival calculus, although the practical applications of subgingival imaging remain limited.

Ridge Preservation After Tooth Extraction With Buccal Bone Plate Deficiency Using Tunnel Structured ß-Tricalcium Phosphate Blocks: A 2-Month Histologic Pilot Study in Beagle Dogs.

BACKGROUND: Reduction in alveolar ridge volume is a direct consequence of tooth extraction. Tunnel ß-tricalcium phosphate (ß-TCP) blocks were manufactured from randomly organized tunnel-shaped ß-TCP ceramic. Efficacy of these blocks compared to extraction alone for alveolar ridge preservation after tooth extraction with buccal bone deficiency was evaluated. METHODS: Maxillary first premolars of six beagle dogs were extracted after removing the buccal bone, and bone defects of 4 × 4 × 5 mm (mesio-distal width × bucco-palatal width × depth) were created. Fresh extraction sockets with buccal bone defects were filled with tunnel ß-TCP blocks at test sites. Two months after the operation, histologic and histometric evaluations were performed. RESULTS: Regarding histologic sections, coronal and middle horizontal widths of the alveolar ridge were significantly greater at test sites (3.2 ± 0.5 and 3.6 ± 0.4 mm, respectively) than at control sites (1.2 ± 0.3 and 2.0 ± 0.6 mm, respectively). The amount of woven bone was significantly greater at test sites (62.4% ± 7.9%) than at control sites (26.8% ± 5.3%), although that of connective tissue and bone marrow was significantly greater at control sites (38.1% ± 6.2% and 16.0% ± 6.9%, respectively) than at test sites (10.7% ± 5.7% and 4.1% ± 2.2%, respectively). Regarding basic multicellular units, no statistically significant difference was found between the test and control sites (0.5% ± 0.1% and 0.6% ± 0.1%, respectively). CONCLUSION: Tunnel ß-TCP blocks represent an effective bone-graft material for alveolar ridge preservation in fresh extraction sockets with buccal bone defects.

Periodontal regeneration using periodontal ligament stem cell-transferred amnion.

Periodontal disease is characterized by the destruction of tooth supporting tissues. Regeneration of periodontal tissues using ex vivo expanded cells has been introduced and studied, although appropriate methodology has not yet been established. We developed a novel cell transplant method for periodontal regeneration using periodontal ligament stem cell (PDLSC)-transferred amniotic membrane (PDLSC-amnion). The aim of this study was to investigate the regenerative potential of PDLSC-amnion in a rat periodontal defect model. Cultured PDLSCs were transferred onto amniotic membranes using a glass substrate treated with polyethylene glycol and photolithography. The properties of PDLSCs were investigated by flow cytometry and in vitro differentiation. PDLSC-amnion was transplanted into surgically created periodontal defects in rat maxillary molars. Periodontal regeneration was evaluated by microcomputed tomography (micro-CT) and histological analysis. PDLSCs showed mesenchymal stem cell-like characteristics such as cell surface marker expression (CD90, CD44, CD73, CD105, CD146, and STRO-1) and trilineage differentiation ability (i.e., into osteoblasts, adipocytes, and chondrocytes). PDLSC-amnion exhibited a single layer of PDLSCs on the amniotic membrane and stability of the sheet even with movement and deformation caused by surgical instruments. We observed that the PDLSC-amnion enhanced periodontal tissue regeneration as determined by micro-CT and histology by 4 weeks after transplantation. These data suggest that PDLSC-amnion has therapeutic potential as a novel cell-based regenerative periodontal therapy.

Periodontal ligament stem cells possess the characteristics of pericytes.

BACKGROUND: Periodontal ligament (PDL) contributes to maintaining homeostasis in periodontal tissues by supplying stem/progenitor cells. It has long been suggested that PDL stem cells/progenitors are located around blood vessels. Recently mesenchymal stem cells (MSCs) have been isolated and cultured from PDL in vitro, although the location of the stem cells in PDL is unclear. The purpose of this study is to test the characteristics of human PDL stem cells (PDLSCs) and examine their similarity to related vascular cell types, such as pericytes and endothelial cells. METHODS: PDLSCs were obtained from healthy extracted teeth using the collagenase/dispase enzyme digestion method. MSC and pericyte characteristics of PDLSCs were examined by cell surface marker expression using flow cytometry. The expression of pericyte markers was tested using immunohistochemistry. Pericyte-like functions of PDLSCs were examined in co-culture of PDLSCs and umbilical vein endothelial cells on a gel matrix. RESULTS: Cultured PDLSCs were positive for both MSC markers and pericyte markers, including cluster of differentiation 146 (CD146), neural/glial antigen 2 (NG2), and CD140b. When pericyte marker expression was explored in rat periodontal tissue sections, CD146- and NG2-positive signals were observed in the perivascular area of the PDL. Further, when the cells were cultured with human umbilical cord endothelial cells under conditions for forming capillary-like structures in vitro, PDLSCs localized adjacent to endothelial cells and contributed to the stability of the capillary-like structure. CONCLUSIONS: PDLSCs possess pericyte-like characteristics and may localize as pericytes in the PDL. These data provide useful information for stem cell biology in periodontal research and stem cell-based periodontal therapy.

Long-term histologic analysis of bone tissue alteration and healing following Er:YAG laser irradiation compared to electrosurgery.

BACKGROUND: The erbium-doped:yttrium, aluminum, and garnet (Er:YAG) laser is reportedly useful for periodontal therapy. However, the potential thermal damage that Er:YAG laser irradiation can produce on bone tissue has not been fully clarified. The purpose of this study was to histologically examine the effects of the Er:YAG laser on bone tissue and subsequent wound healing compared to electrosurgery in a long-term study. METHODS: Calvarial bone from 30 rats was exposed to contact and non-contact Er:YAG laser irradiation (115 mJ/pulse, 10 Hz) without water coolant, or electrode contact. The treated surfaces were analyzed by scanning electron microscopy (SEM), and the healing process was histologically observed until 12 months post-surgery. RESULTS: Contact irradiation resulted in substantial bone ablation, whereas non-contact irradiation produced slight tissue removal. Histologic and SEM analyses of the lased surface showed no severe thermal damage, except for the production of a superficially affected layer with a microstructured surface. The layer did not inhibit new bone formation, and the ablated defect was repaired uneventfully. Although the thickness of the layer gradually decreased, it generally remained in the cortical bone through the observation period. Electrosurgery produced a large area of thermal necrosis without ablation, and the damaged area was not replaced with new bone. CONCLUSIONS: Unlike electrosurgery, Er:YAG laser irradiation without water coolant easily ablated bone tissue, and thermal alteration in the treated surface was minimal. The superficially affected layer did not interfere with the ensuing bone healing, resulting in favorable repair of the defect.

The effect of chemical and/or mechanical conditioning on the Er:YAG laser-treated root cementum: analysis of surface morphology and periodontal ligament fibroblast attachment.

BACKGROUND AND OBJECTIVES: This study compared the surface morphology as well as the biocompatibility of dental root cementum treated with Er:YAG laser irradiation alone and with the laser irradiation followed by chemical and/or mechanical conditioning. STUDY DESIGN/MATERIALS AND METHODS: Healthy cementum plates were randomly assigned to the following control and treatment groups: (1) untreated control (C), (2) Er:YAG laser irradiation (L), (3) laser plus tetracycline HCl (TC) placement (L+TP), (4) laser plus TC burnishing (L+TB), (5) laser plus EDTA gel placement (L+EP), (6) laser plus EDTA gel burnishing (L+EB), (7) laser plus saline solution burnishing (L+SB), and (8) laser plus minocycline-HCl paste placement (L+MP). Specimens were subjected to scanning electron microscopy (SEM), histological observation and attachment assay using periodontal ligament (PDL) fibroblasts. RESULTS: The laser irradiation produced a thin affected layer (5.7 microm thickness) with a superficial microstructure on the cementum surface. The characteristic microstructures of the lased surface were fragile and could be removed by chemical and/or mechanical conditioning treatments. The L+TB group exhibited marked exposure of collagen fibers after removal of the microstructures on the lased surface. The L+EP group presented a peculiar, smooth surface without exposure of collagen fibers and a uniform arrangement of spherical microparticles on the ultra-high magnification of SEM. In cell attachment assay, the L+TB group exhibited the greatest number of attached cells among all the groups, followed by the L+EP, L+SB and control group. The laser alone group exhibited the lowest number of cells. CONCLUSIONS: The characteristic microstructure of the root cementum surface after Er:YAG laser irradiation has a tendency to hinder the early attachment of PDL cells. However, chemical and/or mechanical root conditioning treatment may improve and increase the biocompatibility of the Er:YAG laser-treated root cementum by removing the microstructures of the surface and/or further exposing the collagen fibers.

Er:YAG laser therapy for peri-implant infection: a histological study.

The purpose of this study was to evaluate the effects of Er:YAG laser on degranulation and implant surface debridement in peri-implant infection. The peri-implant infection was experimentally induced in dogs, and the treatment was performed using an Er:YAG laser or a plastic curet. Animals were sacrificed after 24 weeks, and undecalcified histological sections were prepared and analyzed. Degranulation and implant surface debridement were obtained effectively and safely by Er:YAG laser. Histologically, a favorable formation of new bone was observed on the laser-treated implant surface, and the laser group showed a tendency to produce greater bone-to-implant contact than the curet group. These results indicate that the Er:YAG laser therapy has promise in the treatment of peri-implantitis.

Twist negatively regulates osteoblastic differentiation in human periodontal ligament cells.

Periodontal ligament (PDL) is a thin fibrous connective tissue located between two mineralized tissues, alveolar bone and cementum, which maintains a constant width physiologically. The mechanisms by which PDL resists mineralization are not well understood. Twist is a basic helix loop helix protein that plays a central role in regulation of early osteogenesis. We investigated the localization of Twist in PDL and compared the expression of Twist and osteoblast-related genes in PDL cells with those in osteoblast-like cells in the presence or absence of recombinant human bone morphogenetic protein (BMP)-2. Histochemical analysis showed that Twist was expressed along alveolar bone surface in PDL. PDL cells constitutively expressed Twist gene and the expression level was higher than that in osteoblast-like cells. In osteoblast-like cell culture, BMP-2 enhanced osteoblast-related gene expression, while Twist expression was slightly decreased. In contrast, BMP-2 increased runt-related transcription factor (Runx)-2, but failed to enhance alkaline phosphatase (ALP) and osteocalcin (OCN) gene expression in PDL cells. Interestingly, unlike in osteoblast-like cells, Twist expression was upregulated by BMP-2 in PDL cells. We transiently knocked down Twist gene in PDL cells using a short interference RNA expression vector (siTwist) and found that ALP, osteopontin (OPN), bone sialoprotein (BSP) genes expression and basal level of ALP activity were slightly increased, whereas Runx2 and OCN genes were not affected. Collectively, these results suggest that Twist may act as a negative regulator of osteoblastic differentiation in PDL cells.

Antibody response after single-visit full-mouth ultrasonic debridement versus quadrant-wise therapy.

INTRODUCTION: The aim of this study was to compare serum antibody responses to periodontal pathogens after single-visit full-mouth ultrasonic debridement and quadrant-wise therapy. MATERIAL AND METHODS: Thirty-six subjects with chronic periodontitis were randomized into three groups: quadrant-wise debridement in four visits, one-visit full-mouth debridement with water and with povidone iodine. Blood samples were collected before and immediately after treatment and 1, 3 and 6 months post-therapy. Serum antibody titres and avidity to Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Treponema denticola were determined by enzyme-linked immunosorbent assay (ELISA) and thiocyanate ELISA, respectively. RESULTS: IgG titres to P. gingivalis significantly decreased at 1, 3 and 6 months in full-mouth debridement with water group, while significant reductions were seen only at 3 and 6 months after quadrant-wise debridement. Both full-mouth groups showed significant reduction in IgG titres to A. actinomycetemcomitans at 3 and 6 months. Significant increases in antibody avidity to P. gingivalis and A. actinomycetemcomitans were noted 3 months following full-mouth debridement with povidone. CONCLUSION: Both full-mouth and quadrant treatments generally resulted in a decrease in antibody titres and increase in antibody avidity. Full-mouth debridement induced an earlier reduction of IgG titre to P. gingivalis and A. actinomycetemcomitans, than quadrant-wise therapy.

Injectable calcium phosphate bone cement provides favorable space and a scaffold for periodontal regeneration in dogs.

BACKGROUND: An earlier study showed that an injectable calcium phosphate cement (CPC) served as a stable scaffold for bone formation and promoted histocompatible healing of periodontal tissue in dogs. In this study, we evaluated the influence of CPC on regeneration of periodontal defects with experimental periodontitis in dogs. METHODS: Experimental periodontitis was induced by placing stainless-steel mesh on the mesial side of maxillary canines in six adult, healthy beagle dogs. Subsequently, intrabony defects were resized so as to be standard, and CPC was injected in the experimental bone defects. Non-grafted defects on the contralateral side served as controls. Twelve weeks after surgery, the animals were sacrificed and histologic specimens were prepared. Periodontal tissue healing was evaluated histologically and histometrically. RESULTS: Healing of periodontal tissues, in terms of bone and cementum formation, was consistently observed in the CPC-applied sites. CPC was partly replaced by new bone. New cementum and periodontal ligament-like tissue were observed between CPC and the root surface. New bone (P <0.05), new cementum (P <0.01), and new connective tissue attachment and adhesion (P <0.05) were significantly enhanced in the experimental sites. CONCLUSION: Calcium phosphate cement provides stable wound healing and enhanced periodontal regeneration in periodontal defects in dogs with experimental periodontitis.

Periodontal tissue healing following flap surgery using an Er:YAG laser in dogs.

BACKGROUND AND OBJECTIVES: The purpose of this study was to compare periodontal tissue healing following flap surgery using an Er:YAG laser with that of conventional surgery. STUDY DESIGN/MATERIALS AND METHODS: Bilateral premolars with experimentally induced periodontitis in six dogs were treated by periodontal flap surgery. Degranulation and root debridement in the furcation were performed using an Er:YAG laser or curet. At 3 months postsurgery, animals were sacrificed and decalcified specimens were prepared for histological and histometric analysis. RESULTS: Degranulation and root debridement were effectively performed with an Er:YAG laser without major thermal damage and significantly faster than with a curet. Histologically, the amount of newly formed bone was significantly greater in the laser group than in the curet group, although both groups showed similar amounts of cementum formation and connective tissue attachment. CONCLUSIONS: The Er:YAG laser irradiation can be safely and effectively utilized in periodontal flap surgery, and has the potential to promote new bone formation.

Application of periodontal ligament cell sheet for periodontal regeneration: a pilot study in beagle dogs.

OBJECTIVE: The ultimate goal of periodontal treatment is to regenerate the damaged periodontal support. Although periodontal ligament (PDL) cells are essential for periodontal regeneration, few studies have reported the transplantation of periodontal ligament cells to periodontal defects. We developed a new method to apply periodontal ligament cells as a sheet to the defect. The aim of this study was to investigate the periodontal healing after application of the periodontal ligament cell sheet in beagle dogs. METHODS: Autologous periodontal ligament cells were obtained from extracted premolars of each beagle dog. Periodontal ligament cell sheets were fabricated using a temperature-responsive cell culture dish. Dehiscence defects were surgically created on the buccal surface of the mesial roots of bilateral mandibular first molars of each dog. In the experimental group (five defects), periodontal ligament cell sheet with reinforced hyaluronic acid carrier was applied to the defect. Only the hyaluronic acid carrier was applied to the contralateral side as a control (five defects). Eight weeks after surgery, the animals were sacrificed and decalcified specimens were prepared. Healing of the periodontal defects was evaluated histologically and histometrically. RESULTS: No clinical signs of inflammation or recession of gingiva were observed in both experimental and control groups. In the experimental group, periodontal tissue healing with bone, periodontal ligament and cementum formation was observed in three out of five defects. In the control group, such periodontal tissue formation was not observed except in one defect. Histometric analysis revealed that the formation of new cementum in the experimental group was significantly higher than that in the control group. CONCLUSION: The periodontal ligament cell sheet has a potential to regenerate periodontal tissue and may become a novel regenerative therapy.

Effects of the Er:YAG laser irradiation on titanium implant materials and contaminated implant abutment surfaces.

OBJECTIVE: The purpose of this study was to examine the morphological changes and temperature increases of the titanium after Er:YAG laser irradiation, and also to investigate the effect of this laser on debridement of contaminated healing abutments. BACKGROUND DATA: Mechanical instruments have been used for the cleaning of implant abutment surfaces, however, most of them are not appropriate for the application to titanium surface. Recently, the Er:YAG laser has been expected to have a promising ability for the debridement of implant surface. MATERIALS AND METHODS: Experiments were composed of three parts. At first, ten titanium round plates were exposed to the Er:YAG laser irradiation at 30-200 mJ/pulse and the surface changes were observed by stereomicroscope and scanning electron microscope. Secondly, the surface temperature changes of 60 titanium plates during and after Er:YAG laser irradiation at 30 and 50 mJ/pulse were measured by thermographic equipment. At last, calculus on the surface of six contaminated healing abutments was removed by Er:YAG laser or ultrasonic scaler, and the treated surfaces were examined by stereomicroscope. RESULTS: Under 50 mJ/pulse, distinct morphological changes were not observed and the elevation of surface temperature was minimal, especially in the use of water-cooling. The Er:YAG laser at 30 mJ/pulse and 30 Hz with water spray was capable of effectively removing plaque and calculus on the implant abutments without injuring their surfaces. CONCLUSION: This study indicates that the Er:YAG laser can be a novel technical modality for the debridement of implant abutment surface.

Three case reports of aggressive periodontitis associated with Porphyromonas gingivalis in younger patients.

The terms 'early onset periodontitis' (EOP) and 'juvenile periodontitis' (JP) were replaced by that of 'aggressive periodontitis' in a recent international workshop for the classification of periodontal diseases and conditions. The chief etiologic agent for aggressive periodontitis is considered to be Actinobacillus actinomycetemcomitans in localized juvenile periodontitis. Porphyromonas gingivalis is also mentioned as the etiologic agent of the aggressive periodontitis, although to date its role remains questionable. This communication describes three cases of aggressive periodontitis found to be associated with P. gingivalis but not A. actinomycetemcomitans by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Our findings clarify the role of P. gingivalis as an etiologic agent in this type of periodontitis and confirm its inclusion in the current definition of aggressive periodontitis.

Histocompatible healing of periodontal defects after application of an injectable calcium phosphate bone cement. A preliminary study in dogs.

BACKGROUND: A novel injectable, fast setting calcium phosphate cement (CPC) is currently used in orthopedic therapy for bone fractures. This study evaluated the possibility of applying this cement to healing periodontal defects. METHODS: Fenestrations and 3-walled periodontal defects were surgically created on bilateral first molars and canines in 5 beagle dogs. CPC was applied to the defects on one side of the mandible. Untreated defects on the contralateral side served as controls. CPC was applied to all defects in the maxilla. Twelve weeks after surgery, the animals were sacrificed and decalcified and undecalcified specimens were prepared. Periodontal tissue healing was evaluated histologically and histometrically under a light microscope. RESULTS: Healing of periodontal tissues in terms of bone and cementum formation was consistently observed in the CPC-applied sites. CPC was partly replaced by new bone. The residual CPC appeared detached from the denuded root surface. New cementum and periodontal ligament-like tissue were observed between the detached CPC and root surface. No unfavorable reaction was noted in the CPC-applied sites. No statistically significant difference was noted in the experimental or control sites under histometric analysis. CONCLUSIONS: Although there were no statistically significant differences between the 2 treatment groups, histological observation indicated that CPC seemed to act as a scaffold for bone formation and provided histocompatible healing of periodontal tissues in this study. This cement might be applicable to periodontal therapy; however, further investigations are required.