AMPK activation enhances osteoblast differentiation on a titanium disc via autophagy.

PURPOSE: The acquisition of osseointegration during implant therapy is slower and poorer in patients with diabetes compared with healthy persons. The serum concentration of adiponectin in patients with type II diabetes is lower than that of healthy persons via the suppression of AMP-activated protein kinase (AMPK). Therefore, we hypothesized that the AMPK activation enhances bone formation around implants, resulting in the improved acquisition of osseointegration. The purpose of this study was to evaluate the impact of AMPK activation on osteoblast differentiation and its mechanism of downstream signaling on titanium disc (Ti). METHODS: Confluent mouse pre-osteoblasts (MC3T3-E1) cells (1 × 105 cells/well) were cultured with BMP-2 for osteoblast differentiation, in the presence or absence AICAR, an AMPK activator. We examined the effects of AMPK activation on osteoblast differentiation and the underlying mechanism on a Ti using a CCK8 assay, a luciferase assay, quantitative RT-PCR, and western blotting. RESULTS: Although the proliferation rate of osteoblasts was not different between a Ti and a tissue culture polystyrene dish, the addition of AICAR, AMPK activator slightly enhanced osteoblast proliferation on the Ti. AICAR enhanced the BMP-2-dependent transcriptional activity on the Ti, leading to upregulation in the expression of osteogenesis-associated molecules. AICAR simultaneously upregulated the expression of autophagy-associated molecules on the Ti, especially LC3-II. AdipoRon, an adiponectin receptor type1/type2 activator activated AMPK, and upregulated osteogenesis-associated molecules on Ti. CONCLUSIONS: AMPK activation enhances osteoblast differentiation on a Ti via autophagy, suggesting that it promotes the acquisition of osseointegration during implant therapy.

CO2 Laser for Esthetic Healing of Injuries and Surgical Wounds with Small Parenchymal Defects in Oral Soft Tissues.

A number of studies have recently demonstrated the effectiveness of CO2 laser irradiation for the repair and regeneration of scar tissue from injuries or surgical wounds. However, such studies of the oral mucosa are highly limited. Previous studies using CO2 laser irradiation have indicated that two factors contribute to esthetic healing, namely, artificial scabs, which are a coagulated and carbonized blood layer formed on the wound surface, and photobiomodulation therapy (PBMT) for suppressing wound scarring and promoting wound healing. This review outlines basic research and clinical studies of esthetic healing with the use of a CO2 laser for both artificial scab formation by high-intensity laser therapy and PBMT in the treatment of injuries and surgical wounds with small parenchymal defects in oral soft tissues. The results showed that the wound surface was covered by an artificial scab, enabling the accumulation of blood and the perfusion necessary for tissue regeneration and repair. Subsequent PBMT also downregulated the expression of transformation growth factor-b1, which is involved in tissue scarring, and decreased the appearance of myofibroblasts. Taken together, artificial scabs and PBMT using CO2 lasers contribute to the suppression of scarring in the tissue repair process, leading to favorable esthetic and functional outcomes of wound healing.

Development of Novel Decarboxylation-Urea Method toward Interlayer-Anion-Controlled Layered Double Hydroxides.

Layered double hydroxides (LDHs) are representative of a 2D anionic clay. Simple and homogeneous synthesis of interlayer-anion-controlled LDH is essential for studies and industrial production. In this study, we report the one-pot synthesis of an LDH that is selective for interlayer anions, which was labeled as "decarboxylation-urea method". We obtained LDHs intercalated with NO3-, Cl-, and SO42- by removing CO2 in this method. The ionic conductivities of the prepared LDHs were investigated for their applicability to electrolytes, and it was found that Zn-Al LDH intercalated with NO3- showed the highest ionic conductivity (18 mS cm-1). Therefore, the LDH intercalated with NO3- synthesized using the decarboxylation-urea method is promising as an alkaline solid electrolyte.

Current status and factors of periodontal disease among Japanese high school students: a cross-sectional study.

INTRODUCTION: This study aimed to investigate the prevalence of periodontal disease and the factors of the disease among high school students. METHOD: The participants were all students aged 15-18 years (n = 1202) at a high school in Japan. The data on oral health perceptions and behaviours were collected by a questionnaire survey. The prevalence of periodontal disease among them was investigated with the partial community periodontal index (PCPI). A logistic regression analysis was used to identify the factors associated with the PCPI. RESULTS: A total of 1069 students (88.9%) participated in this study. The prevalence of gingival bleeding, calculus, pocket depth of 4-5 mm, and pocket depth of 6 mm or more were 44.2%, 42.2%, 11.4%, and 1.6%, respectively. Approximately one-third of the students had a fear of dental treatment, and only 28.4% used dental floss. The results of logistic regression analysis, adjusted for sex and school year, showed that not visiting dentists regularly, not using dental floss, brushing teeth for less than 5 min, fear of dental treatment, and drinking sports drinks frequently were positively associated with periodontal conditions. CONCLUSION: This study identified a high prevalence of periodontal disease among Japanese high school students aged 15-18 years and its risk factors, such as poor oral health behaviours and fear of dental treatment.

Design of palatal and lingual augmentation prostheses by using an intraoral scanner for a patient after a glossectomy: A clinical report.

Computer-aided design and computer-aided manufacturing was used to fabricate palatal and lingual augmentation prostheses for a patient with dysphagia after a glossectomy. The function of these prostheses was comparable with that of those fabricated by conventional methods. The patient outcome suggests that an intraoral scanner can be effectively used for the fabrication of augmentation prostheses for patients with dysphagia and a high risk of aspiration.

The effect of AMP kinase activation on differentiation and maturation of osteoblast cultured on titanium plate.

Background/purpose: 5' Adenosine monophosphate-activated protein kinase (AMPK) is known as an enzyme that maintains intracellular homeostasis and has various biological activity. The purpose of this study is evaluation effect of AMPK activation on implant prognosis. Materials & methods: MC3T3-E1 osteoblast-like cells were cultured on titanium using a 24-well plate. The experimental group was divided into the following 3 groups: (1) the normal culture group (control group), (2) the osteogenic induction group, and (3) the osteogenic induction + AMPK activation group. The cell counts were measured; real-time PCR was used to assess the expression of ALP and Osterix as osteogenic related genes at Day 0,7,14 and 21 after experiments. Additionally, ALP activity and calcification were assessed. Results: The results of the real-time PCR assessments revealed that the expression of ALP, which is a marker for the initial stages of calcification, was significantly increased by AMPK activation compared to the normal culture or osteogenic induction. A significant increase was also observed in the expression of Osterix, which is a marker for the later stages of calcification. Because significant increases were observed in ALP activity and calcification potential, this suggested that AMPK activation could elicit an increase in osteoblast calcification potential. Conclusion: AMPK activation promotes implant peripheral osteoblast differentiation and maturation and enhances calcification. Our results suggest that AMPK activation may help to maintain implant stability.

Space-making effect for new bone formation by suppressing scar contraction of mucosal epithelium of rat tooth extraction wound using diode laser and CO2 laser treatment.

Background/purpose: As an extraction wound closes, the mucosal epithelium of the tooth extraction wound impedes the space for new bone formation by invading into the extraction socket. Thus, the height of the alveolar crest decreases, causing significant depression of the alveolar mucosa. In this study, we created a rat tooth extraction model and examined the effects of laser irradiation by CO2 and diode on the dynamics of myofibroblast expression through α-SMA, and TGF-ß1. Materials and methods: After tooth extraction of five-week-old male Wistar rats, they were divided into two laser treatment groups (CO2 laser or diode laser was irradiated into tooth extraction socket) and non-laser treatment group (control group). Surrounding tissues, including the extraction socket, were removed at 3, 5, 7, and 21 days after tooth extraction and the expression of α-SMA and TGF-ß1 was verified using immunohistological techniques (6 animals in each group and each period, 72 animals in total). Results: α-SMA-positive cells and TGF-ß1-positive areas were significantly lower in the two laser treatment groups than in the control group. Particularly, the diode group almost had no TGF-ß1-positive areas on the 21st day when healing after tooth extraction was deemed to be completed. Conclusion: Both CO2 and diode laser irradiation of tooth extraction wounds decreases α-SMA-positive cells and TGF-ß1-positive areas. Further, it causes a decrease in myofibroblast expression and suppresses the invasion of mucosal epithelium into the extraction socket. Therefore, laser irradiation may exert a space-making effect for new bone formation and also contribute to socket preservation.

Effects of ytterbium laser surface treatment on the bonding of two resin cements to zirconia.

Monolithic zirconia crowns bonded to zirconia abutments have become more commonly used in the construction of cement-retained implant superstructures. The present study aimed to examine the effects of laser surface treatments on the bond strength of two resin cements to zirconia. Three types of surfaces were examined: untreated, alumina blasted, and ytterbium laser treated; and two types of resin cements: 4-META/MMA-TBB resin cement and composite resin cement. Half of the specimens were subjected to a thermocycling process. Subsequently, a shear bond test was carried out. In addition, surface roughness was measured for each surface type. The results showed that laser treatment increased zirconia surface roughness and that laser treatment significantly increased shear bond strength after the thermocycling of both cement types compared to no treatment. Our experimental results suggested that ytterbium laser surface treatment of zirconia increased the bond strength of resin cements.

Bond durability and surface states of titanium, Ti-6Al-4V alloy, and zirconia for implant materials.

PURPOSE: Screw-retained implant crowns used as dental implants comprise a zirconia coping and titanium base bonded using resin cement. These devices are prone to debonding failures. This study investigated the bond characteristics of implant materials based on shear bond strength (SBS) and surface characteristics. METHODS: Chemically pure (CP) titanium grade-4 (Ti), Ti-6Al-4V alloy (Ti-6Al-4V), and tetragonal polycrystalline zirconia (zirconia) were evaluated as adherent materials. Plates of each material were polished, primed for the respective resin cements, and cemented using either methyl methacrylate-based resin cement (Super-Bond) or composite-based resin cement (Panavia). The cemented samples were subjected to 10,000 thermocycles alternating between 5 and 55 °C, and the SBS were obtained before and after thermocycling. The sample surfaces were characterized based on surface observations, roughness, and free energy (SFE). RESULTS: The SBSs of all materials bonded using Panavia were significantly compromised during thermocycling and reached zero. Although the SBSs of Ti and Ti-6Al-4V bonded using Super-Bond were not significantly affected by thermocycling, those of zirconia decreased significantly. The bond durability between zirconia and Super-Bond was improved via alumina air-abrasion, which caused no significant loss of SBS after thermocycling. Surface analyses of the air-abraded zirconia validated these results and confirmed that its surface roughness and SFE were significantly increased. CONCLUSION: The bond durability between resin cement and zirconia was lower than that between Ti and Ti-6Al-4V. The alumina air-abrasion pretreatment of zirconia improved the SFE and surface roughness, thereby enhancing bond durability.

Late peri-stent contrast staining appearance due to rupture of atherogenic neointima following drug-eluting stent.

The development of peri-stent contrast staining (PSS) after coronary intervention with implantation of a stent is observed in approximately 1-3% of patients treated with drug-eluting stent. Although the cumulative incidences of late in-stent restenosis and stent thrombosis are significantly higher in lesions with PSS than in those without the finding, the mechanisms for the development of PSS have not yet been fully elucidated. In this report, we describe a case of rapid development of PSS with ulcer formation caused by rupture of atherogenic neointima, which was observed by serial optical coherence tomography examinations over 6 months. Protrusion of the stent-jailed underlying necrotic core toward the lumen by the contracting force might have resulted in formation of atherogenic neointima within the stent. Subsequently, rupture of this necrotic core induced by iatrogenic neointimal injury due to balloon dilation and dissolution of the accumulated necrotic core may have resulted in PSS formation 6 months after the procedure. These findings may be helpful for consideration of etiology and therapeutic strategy for lesions with PSS. .

Acute myocardial infarction in a patient with uncorrected tetralogy of Fallot accompanied by coronary artery ectasia: A case report.

A 63-year-old male with a medical history of uncorrected tetralogy of Fallot (TOF) presented to our hospital due to acute myocardial infarction (AMI). Emergency coronary angiography (CAG) was performed and it showed a severe thrombotic stenosis in the middle right coronary artery (RCA) and total thrombotic occlusion of the posterior descending branch of the RCA. Subsequently, percutaneous coronary artery intervention (PCI) under the guidance of intravascular ultrasound (IVUS) was performed. He was discharged on the 14th day in stable condition. Nine months after the PCI procedure, coronary computed tomography angiography was performed for follow-up, which revealed tetralogy of Fallot and complete resolution of the thrombus and ectasic coronary artery without stenosis. When he was 70 years old, he was transferred to our hospital because of recurrent AMI. As emergency CAG showed total thrombotic occlusion of the middle RCA, IVUS-guided PCI was performed. We experienced a very rare case of AMI in an adult patient with uncorrected TOF accompanied by coronary artery ectasia (CAE). To the best of our knowledge, this is the first case of AMI in an adult patient with uncorrected TOF accompanied by CAE. .

An Improved Reflection Colorimeter Integrated with a Coaxial Optical-fiber Cable for Highly Sensitive Solid-phase Colorimetry Using a Membrane Filter.

Highly sensitive solid-phase colorimetry for nickel ion was demonstrated using an improved reflection colorimeter equipped with a coaxial optical-fiber cable. The nickel complex with α-furil dioxime was collected on a small-size membrane filter embedded in a disposable syringe filter unit. The leading edge of the optical-fiber cable was connected to the syringe filter unit via a Luer-lock fitting, and the color intensity of the sample on the filter was evaluated accurately. The detection limit was 0.8 ng in 2.5 mL of the complex solution (0.3 µg L-1). This improved configuration is applicable to highly sensitive on-site analysis without expensive instruments nor high laboratory skills.

Successful percutaneous removal of dislodged ring-marker of optical coherence tomography catheter using the twisted wire technique with a guide-extension catheter: A case report.

An 81-year-old male with diabetes and hypertension was admitted to our hospital due to chest pain on exertion. Coronary angiography revealed a severe stenosis at the middle of right coronary artery (RCA). We performed percutaneous coronary intervention under the guidance of optical coherence tomography (OCT) to the lesion in the middle RCA. After balloon dilations, a drug-eluting stent was deployed to the lesion. Then, OCT examination was performed. At that time, fluoroscopy revealed a foreign body over the 0.014-inch guidewire in the distal RCA, which was the ring-marker of OCT catheter. As RCA blood flow was well preserved, percutaneous removal of the dislodged ring-marker was immediately attempted. At first, we tried to remove the dislodged ring-marker with the guide-extension catheter trapping technique. However, it failed and advanced balloon catheter made the dislodged ring-marker migrate more distally. Therefore, we tried the twisted wire technique with the guide-extension catheter and finally the dislodged ring-marker was removed with it. To the best of our knowledge, this is the first case report of a successful percutaneous removal of a dislodged ring-marker of OCT catheter using the twisted wire technique with a guide-extension catheter. .

Longitudinal Relationship Between Intramuscular Fat in the Quadriceps and Gait Independence in Convalescent Stroke Patients.

BACKGROUND AND AIM: In a cross-sectional study, intramuscular fat in the quadriceps of stroke patients has been associated with gait independence. However, the longitudinal relationship between intramuscular fat and gait independence remain unclear. If these relationships are clarified, it can be demonstrated that improvement in gait independence eventually contributes to improved intramuscular fat in the quadriceps of stroke patients. The aim of this study was to investigate the longitudinal relationship between intramuscular fat in the quadriceps and gait independence in convalescent stroke patients. METHODS: Eleven stroke patients participated in this study. Gait independence was assessed using the Functional Independence Measure (FIM) gait score. The intramuscular fat in the quadriceps was assessed using ultrasound echo intensity, whereas higher echo intensity indicated greater intramuscular fat. The baseline and discharge assessment values for the echo intensity of the quadriceps were compared using a paired t-test. Correlation analysis of the FIM gait score gain and echo intensity changes in the quadriceps on the paretic and non-paretic sides was performed using Kendall's rank correlation coefficient. RESULTS: For quadriceps on the paretic side, echo intensity values at discharge were significantly lower than those at admission. However there was no significant difference for quadriceps on the non-paretic side (paretic side: 19.9% decrease; non-paretic side: 8.0% decrease). We observed that the change in the echo intensity of quadriceps on the non-paretic side was negatively corelated with FIM gait score gain. CONCLUSIONS: Our results revealed a strong correlation between longitudinal change in intramuscular fat in the quadriceps and gait independence, implying that improved gait independence in convalescent stroke patients might have a positive effect on improvements in intramuscular fat.

Effect of DNA/protamine complex paste on bone augmentation of the mandible: A pilot study on dogs.

OBJECTIVE: Our previous studies found that a salmon DNA-based scaffold containing protamine promoted bone regeneration of the calvarial defects of rats. The aim of the present pilot study was to examine the influence of the DNA/protamine (DP) complex on bone regeneration of a saddle type, alveolar ridge defects of the dog mandible. DESIGN: Alveolar ridge defects were performed in the mandibles of five adult female beagles. The following three treatment modalities were randomly allocated: (1) the DP complex paste, (2) a beta-tricalcium phosphate (ß-TCP), and (3) a blank (control). Healing of bone defects were evaluated by periapical radiography, micro-computed tomography (micro-CT), and histology. RESULTS: Periodical radiographic images revealed that a higher percentage of regenerated bone height was consistently achieved in the DP group, as compared with blank controls. All three-dimensional, sagittal, and coronal images of micro-CT showed increased amounts of newly formed bone and a greater bone volume/ tissue volume ratio, as compared with the blank and ß-TCP groups. In contrast, there was no significant difference in bone mineral density among the groups. Histological analysis confirmed that the alveolar bone defects were filled with newly formed bone with mature and compact properties in the DP group. CONCLUSIONS: These findings indicate that the DP complexes enhanced regeneration of vertical alveolar bone defects of the dog mandible.

Effect of neointimal tissue morphology on vascular response to balloon angioplasty in lesions with in-stent restenosis after drug-eluting stent deployment: an optical coherence tomography analysis.

This study aimed to evaluate the vascular response to balloon angioplasty for drug-eluting stent (DES) in-stent restenosis (ISR) lesions based on our novel optical coherence tomography (OCT) classification to establish the optimal treatment strategy for ISR lesions after DES implantation. A total of 104 ISR lesions after DES implantation were imaged by OCT and categorized into the following six patterns: type I-homogeneous high-intensity tissue, type II-heterogeneous tissue with signal attenuation, type III-speckled heterogeneous tissue, type IV-mixed tissue containing poorly delineated region with invisible strut, type V-mixed tissue containing sharply delineated low-intensity region, and type VI-bright protruding tissue with an irregular surface. Serial volumetric OCT analysis was performed before and after balloon dilation to evaluate the vascular response to balloon angioplasty. After balloon dilation, the minimal decrease in neointimal volume was noted in type I lesions and maximal in type III lesions. In contrast, the increase in stent volume was significantly more in type I lesions than others. Neointimal tissue characterization by OCT allows us to provide useful information about the vascular response to balloon dilation, which can influence the therapeutic strategy for DES ISR lesions.

Utility and Validity of Intracoronary Administration of Nicorandil Alone for the Measurement of Fractional Flow Reserve in Patients With Intermediate Coronary Stenosis.

BACKGROUND: Intracoronary (IC) administration of nicorandil has been proposed as an alternative choice of hyperemic agent for fractional flow reserve (FFR) measurements. This study evaluated the utility and validity of IC nicorandil administration alone to induce maximal hyperemia.Methods and Results:Two-hundred-seven patients with coronary artery disease listed for coronary angiography with FFR were prospectively enrolled. FFR was measured after (1) IC administration of nicorandil 2 mg (ICNIC2 mg); (2) continuous intravenous (IV) adenosine triphosphatase (ATP) infusion at 150 µg/kg/min (IVATP150); (3) IV ATP infusion at 210 µg/kg/min (IVATP210); (4) IC administration of 0.5 mg nicorandil during IVATP150 (ICNIC0.5 mg+IVATP150); (5) IC administration of 1 mg nicorandil during IVATP150 (ICNIC1 mg+IVATP150); and (6) IC administration of 2 mg nicorandil during IVATP150 (ICNIC2 mg+IVATP150). The average FFR values and the rate of achieving maximum hyperemia after ICNIC2 mg, IVATP150, IVATP210, ICNIC0.5 mg+IVATP150, ICNIC1 mg+IVATP150, and ICNIC2 mg+IVATP150 were 0.85±0.08, 0.89±0.08, 0.85±0.09, 0.84±0.08, 0.83±0.08, 0.83±0.08 (P<0.01), and 92%, 54%, 91%, 96%, 99%, 99% (P<0.01), respectively. The incidence of systolic aortic pressure drop, chest discomfort, and transient atrioventricular block increased in a dose-dependent manner after IV ATP infusion, but almost no adverse effects were observed after ICNIC2 mg. CONCLUSIONS: ICNIC2 mg produced a more pronounced hyperemia than continuous IV ATP, and might be the preferred method for assessment of FFR.

Accelerated Osteogenic Differentiation and Bone Formation on Zirconia with Surface Grooves Created with Fiber Laser Irradiation.

BACKGROUND: Modification of the surface topography of biomaterials is a critical factor for the proliferation and differentiation of osteoblasts. Elucidating the biological response to surface roughening is necessary for clinical application of zirconia implants. PURPOSE: To investigate the effects of fiber laser-induced zirconia surface roughening on cultured osteoblast-like cell morphology, proliferation, differentiation, and calcification, and on in vivo bone formation. MATERIALS AND METHODS: Sixty-six machine-surfaced yttria-tetragonal zirconia polycrystal plates (S-Zr) and 16 machine-surfaced implants were used as controls. We prepared 66 rough plates (R-Zr) and 16 rough implants by surface treatment using a fiber laser. RESULTS: MC3T3-E1 cells spread well in all directions on S-Zr, whereas elongated cells with poorly organized actin stress fibers were observed on R-Zr. Cell proliferation was significantly greater on R-Zr than on S-Zr. The Runx2 mRNA level increased time dependently in osteogenic culture condition. Alkaline phosphatase activity and osteocalcin mRNA levels were higher on R-Zr compared with S-Zr. Alizarin red S staining revealed greater calcification on R-Zr than on S-Zr. Laser treatment of zirconia implant bodies placed in rat tibiae increased the bone-implant contact ratio and removal torque considerably. CONCLUSIONS: Our results suggest that fiber laser irradiation produces adequate surface roughening of zirconia ceramics to support osseointegration.