The influence on surface characteristic and biocompatibility of nano-SnO2-modified titanium implant material using atomic layer deposition technique.

BACKGROUND/PURPOSE: To investigate the surface characteristics of titanium (Ti) implant materials, which were coated with different thicknesses of nanoscale tin oxide (SnO2) using the atomic layer deposition technique, and evaluated its biological performance on human embryonic palatal mesenchyme (HEPM) cells. METHODS: The thickness of the coating layer on Ti was 0 (Ti0), 20 nm (Ti20), 50 nm (Ti50), and 100 nm (Ti100), respectively. The surface morphology was observed with an SEM and AFM. The root mean square roughness of micron-scale (mRq) and nanoroughness (nRq) of Ti discs' surface were measured. The Alamar blue (AB) assay and F-actin fluorescence staining were used to evaluate the biocompatibility, and the osteocalcin (OCN) was measured to clarify the differentiation of HEPM cells on materials. RESULTS: In the coating groups, the mRq was decreased, but the nRq was increased. The spreading and polygonal morphology of HEPMs was apparent in coating groups. On Day 4, the survival rate of HEPM cells on Ti0 was higher than on Ti20 and Ti50. There was no significant difference on Day 7, Day 10, and Day 14. The OCN was significantly higher on Day 14 in all the coating groups than Ti0. CONCLUSION: The results showed that the cell growth was intensified with rough surfaces. However, the OCN and morphology change was prominent when the nanoroughness was increased, which meant the increased nanoroughness might enhance OCN production and improve the tendency of osseointegration. The nanoscale SnO2 coating could increase the ability of bone formation but not cell growth.

The effects of low-dose 2-hydroxyethyl methacrylate on apoptosis and survival in human dental pulp cells.

BACKGROUND/PURPOSE: 2-hydroxyethyl methacrylate (HEMA) is one of the most major components in dentin bonding systems. Uncured HEMA is eluted through the dentin and harmful to pulp cells. The study aimed to investigate the death pattern, morphological change and factors of human dental pulp cells (HDPCs) cultured with low-dose HEMA. METHODS: HDPCs were cultured with low-dose concentration of HEMA at 0 mM (control), 0.125 mM, 0.25 mM, 1 mM, 2 mM and 4 mM on Day 3 and 5. The cell morphology was observed with F-actin immunocytochemical staining. The flow cytometry was used to analyze the death pattern. NF-κB and Trx-1 were measured using ELISA kits. RESULTS: The major death pattern was early apoptosis and late apoptosis. The morphological characteristics of apoptosis were observed clearly at 4 mM on Day 3 and Day 5. The phosphorylated NF-κB normalized to total NF-κB protein was significantly higher at 2 mM and 4 mM on Day 5. There was no difference of Trx-1 on Day 3, but significantly higher at 0.25 mM and 1 mM on Day 5. The trend line of phosphorylated NF-κB and Trx-1 showed highly positive correlations with HEMA concentration. CONCLUSION: The significant cellular morphology characteristics of apoptosis can be observed at higher dose and longer period after exposed to uncured HEMA. The expression of NF-κB was following the ratio of late apoptosis at longer exposure period. Clinically, the remaining dentin thickness should be enough to decrease HEMA concentration and thus to protect pulp cells free from harm.

Antiplatelet, antioxidative, and anti-inflammatory effects of hydroquinone.

Platelets play crucial roles in thrombosis and hemostasis through platelet activation and aggregation that are crucial in cardiovascular diseases. Hydroquinone (HQ) and its derivatives are present in many dermatological creams, paints, motor fuels, air, microorganisms, and plant products like wheat bread, fruit, coffee, and red wine. The effect of HQ on humans is not clear. In this study, we found that HQ (>25 µM) inhibited arachidonic acid (AA)-induced platelet aggregation. HQ suppressed AA-induced thromboxane B2 production of platelets. HQ (>10 µM) also attenuated ex vivo platelet-rich plasma aggregation. HQ prevented the interleukin (IL)-1ß-induced 8-isoprostane, and PGE2 production, but not IL-8 production of pulp cells. These results indicate that HQ may have an antiplatelet effect via inhibition of thromboxane production. HQ has antioxidative and anti-inflammatory effects, and possible inhibition of COX. Exposure and consumption of HQ-containing products, food or drugs may have antiplatelet, antioxidative, and anti-inflammatory effects.

Prostaglandin E2 Stimulates EP2, Adenylate Cyclase, Phospholipase C, and Intracellular Calcium Release to Mediate Cyclic Adenosine Monophosphate Production in Dental Pulp Cells.

INTRODUCTION: Prostaglandin E2 (PGE2) plays a crucial role in pulpal inflammation and repair. However, its induction of signal transduction pathways is not clear but is crucial for future control of pulpal inflammation. METHODS: Primary dental pulp cells were exposed to PGE2 and 19R-OH PGE2 (EP2 agonist) or sulprostone (EP1/EP3 agonist) for 5 to 40 minutes. Cellular cyclic adenosine monophosphate (cAMP) levels were measured using the enzyme-linked immunosorbent assay. In some experiments, cells were pretreated with SQ22536 (adenylate cyclase inhibitor), H89 (protein kinase A inhibitor), dorsomorphin (adenosine monophosphate-activated protein kinase inhibitor), U73122 (phospholipase C inhibitor), thapsigargin (inhibitor of intracellular calcium release), W7 (calmodulin antagonist), verapamil (L-type calcium channel blocker), and EGTA (extracellular calcium chelator) for 20 minutes before the addition of PGE2. RESULTS: PGE2 and 19R-OH PGE2 (EP2 agonist) stimulated cAMP production, whereas sulprostone (EP1/EP3 agonist) shows little effect. PGE2-induced cAMP production was attenuated by SQ22536 and U73122 but not H89 and dorsomorphin. Intriguingly, thapsigargin and W7 prevented PGE2-induced cAMP production, but verapamil and EGTA showed little effect. CONCLUSIONS: These results indicate that PGE2-induced cAMP production is associated with EP2 receptor and adenylate cyclase activation. These events are mediated by phospholipase C, intracellular calcium release, and calcium-calmodulin signaling. These results are helpful for understanding the role of PGE2 in pulpal inflammation and repair and possible future drug intervention.

IL-1ß-induced MCP-1 expression and secretion of human dental pulp cells is related to TAK1, MEK/ERK, and PI3K/Akt signaling pathways.

OBJECTIVE: Interleukin-1ß (IL-1ß) is an inflammatory molecule of the dental pulp. IL-1ß stimulates cyclooxygenase-2 (COX-2) and prostaglandins production of pulp cells and affects the pulpal inflammation and repair. However, the effects of IL-1ß on Monocyte Chemotactic Factor-1 (MCP-1) of dental pulp cells and its relation to transforming growth factor ß-activated kinase-1 (TAK1), PI3K/Akt, and MEK/ERK signaling and COX activation are not fully clear. DESIGN: Human dental pulp cells were exposed to IL-1ß with/without pretreatment and co-incubation by aspirin (a COX inhibitor), 5z-7-oxozeaenol (a TAK1 inhibitor), LY294002 (a PI3K/Akt inhibitor) or U0126 (a MEK/ERK inhibitor). Viable cell number was evaluated by MTT assay. MCP-1 mRNA expression was tested by reverse transcriptase-polymerase chain reaction (RT-PCR). MCP-1 and COX-2 protein expression was studied by western blot. MCP-1 in the culture medium was measure by ELISA. RESULTS: IL-1ß showed little cytotoxicity to pulp cells. It stimulated MCP-1 mRNA and protein expression and MCP-1 secretion. Aspirin, U0126, LY294002 and 5z-7-oxozeaenol attenuated the IL-1ß-induced MCP-1 expression. In addition, 5z-7-oxozeaenol, LY294002, U0126 and aspirin prevented the IL-1ß-induced MCP-1 secretion of pulp cells. CONCLUSION: These results indicate that IL-1ß may be involved in the pulpal inflammatory and healing processes by inducing MCP-1 expression and secretion. These events are related to differential activation of TAK1, PI3K/Akt and MEK/ERK 1/2 signaling and COX activation. These results are important for future pharmacologic intervention of pulpal inflammatory diseases.

Effects of Camphorquinone on Cytotoxicity, Cell Cycle Regulation and Prostaglandin E2 Production of Dental Pulp Cells: Role of ROS, ATM/Chk2, MEK/ERK and Hemeoxygenase-1.

Camphorquinone (CQ) is a popularly-used photosensitizer in composite resin restoration. In this study, the effects of CQ on cytotoxicity and inflammation-related genes and proteins expression of pulp cells were investigated. The role of reactive oxygen species (ROS), ATM/Chk2/p53 and hemeoxygenase-1 (HO-1) and MEK/ERK signaling was also evaluated. We found that ROS and free radicals may play important role in CQ toxicity. CQ (1 and 2 mM) decreased the viability of pulp cells to about 70% and 50% of control, respectively. CQ also induced G2/M cell cycle arrest and apoptosis of pulp cells. The expression of type I collagen, cdc2, cyclin B, and cdc25C was inhibited, while p21, HO-1 and cyclooxygenase-2 (COX-2) were stimulated by CQ. CQ also activated ATM, Chk2, and p53 phosphorylation and GADD45α expression. Besides, exposure to CQ increased cellular ROS level and 8-isoprostane production. CQ also stimulated COX-2 expression and PGE2 production of pulp cells. The reduction of cell viability caused by CQ can be attenuated by N-acetyl-L-cysteine (NAC), catalase and superoxide dismutase (SOD), but can be promoted by Zinc protoporphyin (ZnPP). CQ stimulated ERK1/2 phosphorylation, and U0126 prevented the CQ-induced COX-2 expression and prostaglandin E2 (PGE2) production. These results indicate that CQ may cause cytotoxicity, cell cycle arrest, apoptosis, and PGE2 production of pulp cells. These events could be due to stimulation of ROS and 8-isoprostane production, ATM/Chk2/p53 signaling, HO-1, COX-2 and p21 expression, as well as the inhibition of cdc2, cdc25C and cyclin B1. These results are important for understanding the role of ROS in pathogenesis of pulp necrosis and pulpal inflammation after clinical composite resin filling.

The effect of implant design and bone quality on insertion torque, resonance frequency analysis, and insertion energy during implant placement in low or low- to medium-density bone.

PURPOSE: This study investigated the effect of implant design and bone quality on insertion torque (IT), implant stability quotient (ISQ), and insertion energy (IE) by monitoring the continuous change in IT and ISQ while implants were inserted in artificial bone blocks that simulate bone of poor or poor-to-medium quality. MATERIALS AND METHODS: Polyurethane foam blocks (Sawbones) of 0.16 g/cm³ and 0.32 g/cm³ were respectively used to simulate low density and low- to medium-density cancellous bone. In addition, some test blocks were laminated with a 1-mm 0.80 g/cm³ polyurethane layer to simulate cancellous bone with a thin cortical layer. Four different implants (Nobel Biocare Mk III-3.75, Mk III-4.0, Mk IV-4.0, and NobelActive-4.3) were placed into the different test blocks in accordance with the manufacturer's instructions. The IT and ISQ were recorded at every 0.5-mm of inserted length during implant insertion, and IE was calculated from the torque curve. The peak IT (PIT), final IT (FIT), IE, and final ISQ values were statistically analyzed. RESULTS: All implants showed increasing ISQ values when the implant was inserted more deeply. In contrast to the ISQ, implants with different designs showed dissimilar IT curve patterns during the insertion. All implants showed a significant increase in the PIT, FIT, IE, and ISQ when the test-block density increased or when the 1-mm laminated layer was present. Tapered implants showed FIT or PIT values of more than 40 Ncm for all of the laminated test blocks and for the nonlaminated test blocks of low to medium density. Parallel-wall implants did not exhibit PIT or FIT values of more than 40 Ncm for all of the test blocks. NobelActive-4.3 showed a significantly higher FIT, but a significantly lower IE, than Mk IV-4.0. CONCLUSIONS: While the existence of cortical bone or implant designs significantly affects the dynamic IT profiles during implant insertion, it does not affect the ISQ to a similar extent. Certain implant designs are more suitable than others if high IT is required in bone of poor quality. The manner in which IT, IE, and ISQ represent the implant primary stability requires further study.

Comparison of clinical parameters and environmental noise levels between regular surgery and piezosurgery for extraction of impacted third molars.

BACKGROUND/PURPOSE: Impacted third molars can be extracted by regular surgery or piezosurgery. The aim of this study was to compare clinical parameters and device-produced noise levels between regular surgery and piezosurgery for the extraction of impacted third molars. METHODS: Twenty patients (18 women and 2 men, 17-29 years of age) with bilateral symmetrical impacted mandibular or maxillary third molars of the same level were included in this randomized crossover clinical trial. The 40 impacted third molars were divided into a control group (n = 20), in which the third molar was extracted by regular surgery using a high-speed handpiece and an elevator, and an experimental group (n = 20), in which the third molar was extracted by piezosurgery using a high-speed handpiece and a piezotome. The clinical parameters were evaluated by a self-reported questionnaire. The noise levels produced by the high-speed handpiece and piezotome were measured and compared between the experimental and control groups. RESULTS: Patients in the experimental group had a better feeling about tooth extraction and force delivery during extraction and less facial swelling than patients in the control group. However, there were no significant differences in noise-related disturbance, extraction period, degree of facial swelling, pain score, pain duration, any noise levels produced by the devices under different circumstances during tooth extraction between the control and experimental groups. CONCLUSION: The piezosurgery device produced noise levels similar to or lower than those of the high-speed drilling device. However, piezosurgery provides advantages of increased patient comfort during extraction of impacted third molars.

Simvastatin suppresses osteoblastic expression of Cyr61 and progression of apical periodontitis through enhancement of the transcription factor Forkhead/winged helix box protein O3a.

INTRODUCTION: In this study, the role of transcription factor Forkhead/winged helix box protein O3a (FoxO3a) in Cyr61 expression and its modulation by simvastatin were investigated in cultured murine osteoblasts and a rat model of induced apical periodontitis. We also examined the effects of simvastatin on the synthesis of chemokine CCL2 and chemotaxis of macrophages in vitro. METHODS: We assessed tumor necrosis factor (TNF)-α-stimulated expression of Cyr61 and phosphorylated inactive FoxO3a (p-FoxO3a) in MC3T3-E1 murine osteoblasts by Western analysis. Forced expression of FoxO3a by lentiviral-based gene transduction was performed, and its effect on Cyr61 expression was evaluated. The modulation of CCL2 secretion and macrophage chemotaxis by simvastatin were examined by enzyme-linked immunosorbent assay and transwell migration assay, respectively. In a rat model of induced apical periodontitis, the relation between disease progression and osteoblastic expression of Cyr61, p-FoxO3a, and CCL2 and macrophage recruitment were studied by radiographic and immunohistochemistry analyses. RESULTS: Western blot analysis showed enhanced expression of Cyr61 and p-FoxO3a after TNF-α treatment in a time-dependent manner. Simvastatin significantly counteracted the actions of TNF-α. Forced expression of FoxO3a reduced TNF-α-stimulated Cyr61 synthesis. Simvastatin and FoxO3a diminished TNF-α-induced CCL2 secretion and macrophage recruitment, whereas Cyr61 partially restored the stimulating action. In rat periapical lesions, simvastatin significantly attenuated bone resorption, reduced osteoblastic expressions of Cyr61, p-FoxO3a, and CCL2, and suppressed macrophage recruitment. CONCLUSIONS: Simvastatin may alleviate periapical lesions by enhancing FoxO3a activity to suppress the synthesis of Cyr61 in osteoblasts. Moreover, the downstream effector mechanism of Cyr61 may involve CCL2 production and macrophage recruitment.

Monomer conversion and cytotoxicity of dental composites irradiated with different modes of photoactivated curing.

The aim of the study was to compare the monomer conversion and cytotoxicity of two resin composites, Tetric Ceram (TC) and Heliomolar (HM), irradiated with a halogen light-curing unit (Optilux 501) using a number of curing modes, boost (OB), ramp (OR), and conventional (OC), as well as a light-emitting diode (LED) curing unit (LEDemetron; LEDe). The effects of irradiation times (10, 20, 30, and 40 s) were also investigated. The monomer conversion of resin composites was measured using Fourier transform infrared (FTIR) analysis. Resin samples were preimmersed in Dulbeco's modified engles medium (DMEM) for 24 h, and the periodontal ligament cells (PDL) were cultured with aged medium for 72 h to evaluate the cytotoxic effects of resin samples using the microtiter-tetrazolium (MTT) method. The spectral outputs of the curing units were also compared. After irradiation using the same curing mode, the TC monomer conversion was higher than the HM analog. The TC conversion value was lower after OC and LEDemetron irradiation for 10 s compared to exposures of 30 and 40 s. The HM conversion value was lowest for the 10-s OC irradiation, and highest for the 40-s OC exposure. In cytotoxicity tests, the negative control group was without irradiation. The succinate dehydrogenase (SDH) activities of the OB and OR-irradiated TC samples were significantly lower than analogs irradiated with LEDemetron for 40 s; however, no significant differences were demonstrated between OB, OR, and control groups. Further, no significant differences in SDH activity were demonstrated for OB, OR, and control groups of the HM.

Effects on microstrain and conversion of flowable resin composite using different curing modes and units.

The flowable resin composite, Tetric Flow, was used to measure microstrain and degree of conversion after hardening with each of three curing machines: XL3000(XL) for 10, 20, 30, and 40 s; Optilux 501 using conventional mode (OC) for 10, 20, 30, and 40 s, as well as Optilux boost (OB, 10 s) and ramp modes (OR, 20 s); and LEDemetron (LEDe) for 10, 20, 30, and 40 s. The emitted power density and spectral distribution of the three light curing units were also measured. The LEDe output energy spectrum was centralized between 425 and 490 nm, which encompasses the excited wavelength of camphorquinone. The microstrain produced by the curing process is as a second-degree polynomial for each light source. The OB microstrain was highest, while the OR microstrain was lower. The ranking in order of degree of monomer conversion was as follows: XL10

Intrusion and reversal of a free-standing natural tooth bounded by two implant-supported prostheses: a clinical report.

Although intrusion of natural tooth abutments in tooth-implant connected fixed prostheses has been reported, it can also occur to a free-standing natural tooth bounded by implant prostheses. For the patient described in this article, intrusion was noted with a natural tooth bounded by 2 implant-supported prostheses 5 months after insertion of the prostheses. The intrusion was reversed completely after 5 months with appropriate management. The course of treatment and possible mechanisms of intrusion are provided.

The role of lipopolysaccharide in infectious bone resorption of periapical lesion.

BACKGROUND: The role of lipopolysaccharide (LPS) in periapical lesion-induced bone resorption was investigated. Polymyxin B (PMB), a specific inhibitor of LPS, was evaluated to treat the apical lesion. METHODS: Lipopolysaccharide isolated from two common endodontic pathogens, Fusobacterium nucleatum and Porphyromonas endodontalis, stimulated mouse macrophage (J774) to release interleukin-1alpha (IL-1 alpha) and tumor necrosis factor-alpha (TNF-alpha) in a time-dependent manner. RESULTS: Combination of LPS further enhanced the stimulation. PMB inhibited these effects significantly. LPS also stimulated matrix metalloproteinase-1 (MMP-1) gene expression in J774, whereas anti-IL-1 alpha and anti-TNF-alpha antibodies, as well as PMB, diminished this effect. A disease model of periapical lesion was established in Wistar rat. Administration of PMB reduced the extent of lesion-associated bone resorption by 76% to approximately 80%, and simultaneously reduced the numbers of MMP-1-producing macrophages. CONCLUSIONS: It is suggested that LPS released from the infected root canal triggers the synthesis of IL-1 alpha and TNF-alpha from macrophages. These pro-inflammatory cytokines up-regulate the production of MMP-1 by macrophages to promote periapical bone resorption.