Effect of photodynamic therapy according to differences in photosensitizers on Staphylococcus aureus biofilm on titanium.

PURPOSE: This study aimed to evaluate the antimicrobial effect of photodynamic therapy (PDT) against Staphylococcus aureus biofilm on a titanium surface and to compare the differences in the effect of PDT using toluidine blue O (TBO) and methylene blue (MB) as a photosensitizer. METHODS: The bacterial strain S. aureus ATCC 25,923 was used. Sandblasted and acid-etched (SLA) disks were divided into the following six groups: phosphate buffer saline (PBS), TBO, MB, PBS with laser (PBS + L), TBO with laser (TBO + L), and MB with laser (MB + L). The laser group samples were irradiated by a cold diode laser for 60 s. After treatment, the number of surviving bacteria was calculated by counting the colony-forming units (CFUs) and confocal laser scanning microscopy (CLSM) was applied to observe the bacteria on the disk surface. RESULTS: The TBO + L and MB + L groups showed significantly lower CFU/ml than the other groups (p < 0.01). The TBO + L group showed significantly lower CFU/ml than the MB + L group (p = 0.032). There was no significant difference between the PBS, TBO, MB, and PBS + L groups. Within the limitations of this in vitro study, PDT with TBO and MB can effectively reduce S. aureus biofilm on SLA titanium surfaces. TBO is more effective than MB as a photosensitizer. PDT with TBO may be applied to the treatment of peri­implant disease in the future.

Temporal changes of periodontal tissue pathology in a periodontitis animal model.

PURPOSE: This study aimed to characterize the early stages of periodontal disease and determine the optimal period for its evaluation in a mouse model. The association between the duration of ligation and its effect on the dentogingival area in mice was evaluated using micro-computed tomography (CT) and histological analysis. METHODS: Ninety mice were allocated to an untreated control group or a ligation group in which periodontitis was induced by a 6-0 silk ligation around the left second maxillary molar. Mice were sacrificed at 1, 2, 3, 4, 5, 8, 11, and 14 days after ligature placement. Alveolar bone destruction was evaluated using micro-CT. Histological analysis was performed to assess the immune-inflammatory processes in the periodontal tissue. RESULTS: No significant difference in alveolar bone loss was found compared to the control group until day 3 after ligature placement, and a gradual increase in alveolar bone loss was observed from 4 to 8 days following ligature placement. No significant between-group differences were observed after 8 days. The histological analysis demonstrated that the inflammatory response was evident from day 4. CONCLUSIONS: Our findings in a mouse model provide experimental evidence that ligature-induced periodontitis models offer a consistent progression of disease with marginal attachment down-growth, inflammatory infiltration, and alveolar bone loss.

Long-term effect of implant-abutment connection type on marginal bone loss and survival of dental implants.

PURPOSE: This study aimed to compare the long-term survival rate and peri-implant marginal bone loss between different types of dental implant-abutment connections. METHODS: Implants with external or internal abutment connections, which were fitted at Gangneung-Wonju National University Dental Hospital from November 2011 to December 2015 and followed up for >5 years, were retrospectively investigated. Cumulative survival rates were evaluated for >5 years, and peri-implant marginal bone loss was evaluated at 1- and 5-year follow-up examinations after functional loading. RESULTS: The 8-year cumulative survival rates were 93.3% and 90.7% in the external and internal connection types, respectively (P=0.353). The mean values of marginal bone loss were 1.23 mm (external) and 0.72 mm (internal) (P<0.001) after 1 year of loading, and 1.20 mm and 1.00 mm for external and internal abutment connections, respectively (P=0.137) after 5 years. Implant length (longer, P=0.018), smoking status (heavy, P=0.001), and prosthetic type (bridge, P=0.004) were associated with significantly greater marginal bone loss, and the use of screw-cement-retained prosthesis was significantly associated (P=0.027) with less marginal bone loss. CONCLUSIONS: There was no significant difference in the cumulative survival rate between implants with external and internal abutment connections. After 1 year of loading, marginal bone loss was greater around the implants with an external abutment connection. However, no significant difference between the external and internal connection groups was found after 5 years. Both types of abutment connections are viable treatment options for the reconstruction of partially edentulous ridges.

Antimicrobial photothermal therapy using diode laser with indocyanine green on Streptococcus gordonii biofilm attached to zirconia surface.

PURPOSE: The purpose of this study was to evaluate the antimicrobial effects of photothermal therapy using indocyanine green (ICG) and an 810-nm infrared diode laser on Streptococcus gordonii biofilm attached to zirconia surfaces in vitro. METHODS: A biofilm was formed using the static method on zirconia disks placed in a 24-well plate. The biofilms were subdivided into the following six treatment groups: control, commercial photodynamic therapy (PDT), chlorhexidine gluconate (CHX), laser only (L, 810-nm infrared diode), ICG, and laser with ICG (PTT). After treatment, each disk was agitated and the solution with detached bacteria was spread directly on a blood agar plate. Cells were cultured under anaerobic conditions and colony-forming units were counted. Confocal laser-scanning microscopy was used to assess the survival according to the height of the biofilm. RESULTS: The PTT, PDT, and CHX groups showed a significant reduction in S. gordonii viability (p<0.05), while the L and ICG groups showed no significant difference compared to the control group (p = 0.32, p = 0.97; respectively). In confocal laser-scanning microscopy images, the PTT, PDT, and CHX groups presented most of the dead bacteria in both the upper and lower levels of biofilm. CONCLUSION: Within the limitations of this in vitro study, PTT with ICG was effective in significantly reducing the viability of S. gordonii bacteria on zirconia. Further studies are needed to establish a standardized PTT protocol to treat peri­implant diseases.

Effects of ibuprofen-loaded TiO2 nanotube dental implants in alloxan-induced diabetic rabbits.

PURPOSE: Some systemic conditions, especially diabetes mellitus (DM), adversely affect dental implant success. This study aimed to investigate the effects of ibuprofen-loaded TiO2 nanotube (ILTN) dental implants in alloxan-induced diabetic rabbits. METHODS: Twenty-six New Zealand white rabbits were treated with alloxan monohydrate to induce DM. At 2 weeks following DM induction, 3 types of implants (sandblasted, large-grit, and acid-etched [SLA], ILTN, and machined) were placed into the proximal tibia in the 10 rabbits that survived following DM induction. Each type of implant was fitted randomly in 1 of the holes (round-robin method). The animals were administered alizarin (at 3 weeks) and calcein (at 6 weeks) as fluorescent bone markers, and were sacrificed at 8 weeks for radiographic and histomorphometric analyses. RESULTS: TiO2 nanotube arrays of ~70 nm in diameter and ~17 µm in thickness were obtained, and ibuprofen was loaded into the TiO2 nanotube arrays. A total of 26 rabbits were treated with alloxan monohydrate and only 10 rabbits survived. The 10 surviving rabbits showed a blood glucose level of 300 mg/dL or higher, and the implants were placed in these diabetic rabbits. The implant stability quotient (ISQ) and bone-to-implant contact (BIC) values were significantly higher in the ILTN group (ISQ: 61.8, BIC: 41.3%) and SLA group (ISQ: 62.6, BIC: 46.3%) than in the machined group (ISQ: 53.4, BIC: 20.2%), but the difference in the BIC percentage between the SLA and ILTN groups was not statistically significant (P=0.628). However, the bone area percentage was significantly higher in the ILTN group (78.0%) than in the SLA group (52.1%; P=0.000). CONCLUSIONS: The ILTN dental implants showed better stability (ISQ) and BIC than the machined implants; however, these values were similar to the commercially used SLA implants in the 2-week diabetic rabbit model.

Clinical and microbiological effects of adjunctive local delivery of minocycline (Periocline®) in patients receiving supportive periodontal therapy: a pilot study.

PURPOSE: This study aimed to evaluate the clinical and microbiological efficacy of adjunctive local delivery of minocycline (Periocline®) in patients receiving supportive periodontal therapy (SPT) after initial treatment. METHODS: The participants were 16 men and 8 women (age, 20-65 years) who had at least 15 natural teeth, underwent SPT for more than 1 year due to chronic periodontitis, had 4 or more periodontal pocket sites deeper than 5 mm, and showed >25% gingival bleeding on probing (BoP). They were randomly assigned to the test and control groups. In the test group, mechanical debridement and local antibiotic delivery were performed for all periodontal sulci/pockets; in the control group, mechanical debridement and saline irrigation were performed. In patients who underwent SPT for more than 1 year, clinical and microbiological examinations were performed at baseline and 1 and 3 months after SPT. The clinical examination included an assessment of the periodontal pocket depth, clinical attachment level, plaque index, and BoP. Microbial tests were performed using real-time polymerase chain reaction; the relative ratios of Porphyromonas gingivalis and Fusobacterium nucleatum were determined. RESULTS: Both groups showed significant improvements in clinical parameters at 1 and 3 months from baseline; there were no significant changes between months 1 and 3. Intergroup differences were insignificant. The microbiological analysis revealed no significant differences in P. gingivalis and F. nucleatum ratios across time points. While intergroup differences were insignificant, there was a tendency for the P. gingivalis and F. nucleatum ratios to decrease in the test group. CONCLUSIONS: Mechanical debridement in patients receiving maintenance therapy resulted in clinically significant improvement; the effectiveness of additional local delivery of antibiotics was not significant. The ratios of P. gingivalis and F. nucleatum showed a tendency to decrease in the test group, although it was not significant.

Photofunctionalizing effects of hydroxyapatite combined with TiO2 on bone regeneration in rabbit calvarial defects.

The hydrophilicity of bone graft material generally used as a carrier can play an important role in regulating bone morphogenetic protein (BMP) expression at the bone graft site. The hydrophilicity, altering physicochemical properties, and enhancing biological capabilities, can be increased via surface modification through ultraviolet (UV) photofunctionalization and the effect on de novo osteogenesis could be further improved. Therefore, this study aimed to assess the effects of UV-irradiated TiO2 -coated hydroxyapatite (HA) in combination with rhBMP-2 on bone regeneration in rabbit calvarial defects. The hydrophilicity of HA and TiO2 -coated HA pellets was evaluated by measuring the contact angle of water droplets with UV irradiation. To compare de novo osteogenesis in rabbit calvarial defects, the rabbits were segregated into four different groups: negative control, HA, TiO2 -coated HA, and TiO2 -coated HA with UV; histomorphometric analysis and micro-computed tomography (µCT) imaging were performed after 4 and 8 weeks. In vivo analysis revealed that de novo osteogenesis occurred on the critical size defects in all groups and was significantly increased in the TiO2 -coated HA with UV group than in other groups (p < 0.05). The present results indicate that UV photofunctionalization promotes de novo osteogenesis. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1953-1959, 2019.

Efficacy of an LED toothbrush on a Porphyromonas gingivalis biofilm on a sandblasted and acid-etched titanium surface: an in vitro study.

PURPOSE: The aim of this study was to evaluate the antimicrobial effect of a newly devised toothbrush with light-emitting diodes (LEDs) on Porphyromonas gingivalis attached to sandblasted and acid-etched titanium surfaces. METHODS: The study included a control group, a commercial photodynamic therapy (PDT) group, and 3 test groups (B, BL, and BLE). The disks in the PDT group were placed in methylene blue and then irradiated with a diode laser. The B disks were only brushed, the BL disks were brushed with an LED toothbrush, and the BLE disks were placed into erythrosine and then brushed with an LED toothbrush. After the different treatments, bacteria were detached from the disks and spread on selective agar. The number of viable bacteria and percentage of bacterial reduction were determined from colony counts. Scanning electron microscopy was performed to visualize bacterial alterations. RESULTS: The number of viable bacteria in the BLE group was significantly lower than that in the other groups (P<0.05). Scanning electron microscopy showed that bacterial cell walls were intact in the control and B groups, but changed after commercial PDT and LED exposure. CONCLUSIONS: The findings suggest that an LED toothbrush with erythrosine treatment was more effective than a commercial PDT kit in reducing the number of P. gingivalis cells attached to surface-modified titanium in vitro.

Determination of the optimal diabetes duration for bone regeneration experiments in an alloxan-induced diabetic rabbit calvarial defect model.

PURPOSE: The purpose of this study was to evaluate the optimal diabetes duration for bone regeneration experiments in an alloxan monohydrate (ALX)-induced diabetic rabbit calvarial defect model by evaluating the association between diabetes duration and bone healing capacity. METHODS: Twenty-four New Zealand white rabbits were used. Twenty-two rabbits were injected with 100 mg/kg of ALX to induce experimental diabetes. These rabbits were divided into 4 groups, including a control group and groups with diabetes durations of 1 week (group 1), 2 weeks (group 2), and 4 weeks (group 3). Calvarial defects were created at 1, 2, and 4 weeks after ALX injection and in the control rabbits. Cone-beam computed tomography (CBCT) scanning was performed on the day of surgery and at 2 and 4 weeks after surgery. The rabbits were sacrificed 4 weeks after surgery, followed by histological and immunofluorescence analysis. RESULTS: The diabetic state of all diabetic rabbits was well-maintained throughout the experiment. Reconstructed 3-dimensional CBCT imaging showed more rapid and prominent bone regeneration in the control group than in the experimental groups. Histological staining showed notable bone regeneration in the control group, in contrast to scarce bone formation in the experimental groups. The appearance and immunoreactivity of receptor activator of nuclear factor-kappa B and osteoprotegerin did not show notable differences among the groups. CONCLUSION: ALX administration at 100 mg/kg successfully induced experimental diabetes in rabbits. The effect of diabetes on bone healing was evident when the interval between diabetes induction and the intervention was ≥1 week.

Determination of the critical diabetes duration in a streptozotocin-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration.

PURPOSE: The purpose of this study was to determine the critical diabetes duration in a streptozotocin (STZ)-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration by evaluating the association between diabetes duration and bone healing capacity through histological and radiographic analyses. METHODS: Experimental diabetes was induced in 50 of 60 rats by an STZ injection. The rats were divided into 5 groups, including a control group (group 1), according to diabetes durations of 0, 2, 4, 6, and 8 weeks, respectively. Eighteen rats survived: 4 in group 1, 4 in group 2, 4 in group 3, 5 in group 4, and 1 in group 5. Calvarial defects were created at 0, 2, 4, 6, and 8 weeks after STZ injection in groups 1-5. Cone-beam computed tomography scanning was performed at baseline and at 5 and 7 weeks after surgery. The rats were sacrificed 7 weeks after surgery, followed by histological evaluation. RESULTS: The voxel gray values (VGVs) of group 1 and group 2 increased, whereas the VGVs of group 3 and group 4 decreased starting 5 weeks after surgery, although this trend did not reach statistical significance between groups. On the reconstructed 3-dimensional images and based on an analysis of histological features, groups 1 and 2 showed apparent bone regeneration, while groups 3-5 showed very limited bone regeneration. CONCLUSIONS: The critical diabetes duration in an STZ-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration was between 2 and 4 weeks. It is suggested that researchers who use STZ-induced diabetic rats wait for more than 2 weeks following diabetes induction before placing implants or conducting bone regeneration studies to allow definite disturbances in bone healing to emerge.

Comparison of periodontitis-associated oral biofilm formation under dynamic and static conditions.

PURPOSE: The purpose of this study was to compare the characteristics of single- and dual-species in vitro oral biofilms made by static and dynamic methods. METHODS: Hydroxyapatite (HA) disks, 12.7 mm in diameter and 3 mm thick, were coated with processed saliva for 4 hours. The disks were divided into a static method group and a dynamic method group. The disks treated with a static method were cultured in 12-well plates, and the disks in the dynamic method group were cultured in a Center for Disease Control and Prevention (CDC) biofilm reactor for 72 hours. In the single- and dual-species biofilms, Fusobacterium nucleatum and Porphyromonas gingivalis were used, and the amount of adhering bacteria, proportions of species, and bacterial reduction of chlorhexidine were examined. Bacterial adhesion was examined with scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). RESULTS: Compared with the biofilms made using the static method, the biofilms made using the dynamic method had significantly lower amounts of adhering and looser bacterial accumulation in SEM and CLSM images. The proportion of P. gingivalis was higher in the dynamic method group than in the static method group; however, the difference was not statistically significant. Furthermore, the biofilm thickness and bacterial reduction by chlorhexidine showed no significant differences between the 2 methods. CONCLUSIONS: When used to reproduce periodontal biofilms composed of F. nucleatum and P. gingivalis, the dynamic method (CDC biofilm reactor) formed looser biofilms containing fewer bacteria than the well plate. However, this difference did not influence the thickness of the biofilms or the activity of chlorhexidine. Therefore, both methods are useful for mimicking periodontitis-associated oral biofilms.

The antiplaque and bleeding control effects of a cetylpyridinium chloride and tranexamic acid mouth rinse in patients with gingivitis.

PURPOSE: This study aimed to evaluate the effects of a cetylpyridinium chloride (CPC) and tranexamic acid (TXA) mouth rinse on patients with gingivitis. METHODS: This randomized, placebo-controlled, double-blind, parallel-group, clinical trial included 45 healthy adults with gingivitis, who were randomized into 2 groups. The experimental group used a 0.05% CPC and 0.05% TXA mouth rinse, and the control group used a placebo mouth rinse. The following clinical indices were assessed at baseline, at 3 weeks, and at 6 weeks: the Turesky-Quigley-Hein plaque index (QHI), the Löe-Silness gingival index (GI), and bleeding on marginal probing (BOMP). The subjects used the mouth rinse during the experimental period for 20 seconds, 4-5 times daily (10 mL each time). RESULTS: There were no significant differences in the clinical indices between the groups at baseline. In the experimental group (CPC+TXA), a statistically significant improvement was evident in the QHI, GI, and BOMP at 3 and 6 weeks. These results were similar to those observed in the control group at 3 and 6 weeks, although the change in BOMP was not statistically significant in that group. At 6 weeks, the experimental group had a significantly lower mean score for the QHI than the control group. CONCLUSIONS: This study demonstrated that a CPC and TXA mouth rinse exhibited significant antiplaque and anti-gingivitis efficacy, and had a positive effect on bleeding control when used daily for 6 weeks.

Serotonin Inhibits Osteoblast Differentiation and Bone Regeneration in Rats.

BACKGROUND: Although increasing evidence indicates that serotonin (SER; 5-hydroxytrypamine [5-HT]) is involved in the regulation of bone metabolism, conflicting data exist regarding whether SER promotes or inhibits osteoblast differentiation and bone formation. Regeneration of functional bone is required for proper osseointegration of dental implants. Noticeably, the use of selective SER reuptake inhibitors was recently associated with the failure of osseointegrated dental implants. The present study examines the direct role of peripheral SER on the regulation of bone regeneration. METHODS: The effect of SER on osteoblast differentiation and bone regeneration was examined using rat calvarial cell cultures in vitro and a rat critical-sized calvarial defect model in vivo. RESULTS: Rat calvarial cells expressed SER receptors Htr1 (5-HT1) and Htr2 (5-HT2), which are known to transmit signals in bone cells. In vitro, SER significantly reduced osteogenic differentiation and mineralization of rat calvarial cells with concomitant reduction of osteoblast marker genes including alkaline phosphatase (Alpl), osterix (Sp7), and osteocalcin (Bglap). Histologic and radiologic analyses using the rat critical-sized calvarial defect model revealed that the existence of SER significantly inhibited ß-phase tricalcium phosphate-induced bone regeneration. CONCLUSION: Results suggest that SER in the local bone microenvironment might play a negative role in osteoblast differentiation and bone formation in rats.

The effect of photodynamic therapy on Aggregatibacter actinomycetemcomitans attached to surface-modified titanium.

PURPOSE: The purpose of this study was to evaluate the effect of photodynamic therapy (PDT) using erythrosine and a green light emitting diode (LED) light source on biofilms of Aggregatibacter actinomycetemcomitans attached to resorbable blasted media (RBM) and sandblasted, large-grit, acid-etched (SLA) titanium surfaces in vitro. METHODS: RBM and SLA disks were subdivided into four groups, including one control group and three test groups (referred to as E0, E30, E60), in order to evaluate the effect of PDT on each surface. The E0 group was put into 500 µL of 20 µM erythrosine for 60 seconds without irradiation, the E30 group was put into erythrosine for 60 seconds and was then irradiated with a LED for 30 seconds, and the E60 group was put into erythrosine for 60 seconds and then irradiated with a LED for 60 seconds. After PDT, sonication was performed in order to detach the bacteria, the plates were incubated under anaerobic conditions on brucella blood agar plates for 72 hours at 37℃, and the number of colony-forming units (CFUs) was determined. RESULTS: Significant differences were found between the control group and the E30 and E60 groups (P<0.05). A significantly lower quantity of CFU/mL was found in the E30 and E60 groups on both titanium disk surfaces. In confocal scanning laser microscopy images, increased bacterial death was observed when disks were irradiated for a longer period of time. CONCLUSIONS: These findings suggest that PDT using erythrosine and a green LED is effective in reducing the viability of A. actinomycetemcomitans attached to surface-modified titanium in vitro.

IL-17 inhibits osteoblast differentiation and bone regeneration in rat.

OBJECTIVE: The interleukin-17 (IL-17) family is a group of pro-inflammatory cytokines that are produced by a subset of helper T cells. IL-17 family members are not only involved in the immune response of tissues but also play a role in bone metabolism. Although the role of IL-17 in osteoclast-mediated bone resorption has been extensively studied, its role during osteoblast-mediated bone formation has rarely been investigated. In this study, we examined the effect of IL-17 on osteogenesis in rats both in vitro and in vivo. DESIGN: To evaluate osteogenesis in vitro, rat calvarial osteoblast precursor cells were cultured for 14 days in osteogenic medium with or without 50ng/mL IL-17. Osteogenic activity was evaluated by alkaline phosphatase and alizarin red staining. The mRNA expression of alkaline phosphatase, osteocalcin, and osterix was also measured by using real-time PCR. To test whether IL-17 affects bone formation in vivo, bone filling was examined by micro-computed tomography and histological observations at 8 weeks after critical-sized defects were made in rat calvaria. RESULTS: The presence of IL-17 significantly reduced alkaline phosphatase and alizarin red staining and the expression of alkaline phosphatase, osteocalcin, and osterix in vitro. IL-17 also significantly inhibited the filling of calvarial defects in vivo. CONCLUSION: IL-17 exerted a negative effect on osteogenesis in a rat model. In contrast to the previously reported beneficial effect on osteogenic differentiation of human mesenchymal stem cells, our results suggest a species or cell type-specific role for IL-17 in bone formation.

Effect of supportive periodontal therapy on the prevention of tooth loss in Korean adults.

PURPOSE: The purpose of this retrospective study was to evaluate the effect of patient compliance with supportive periodontal therapy (SPT) on tooth loss in Korean adults. METHODS: The periodontal records of 134 patients were reviewed for this study. They completed active periodontal treatment from 1999 to 2001 and were placed on a schedule of periodic follow-up visits for SPT. Patient compliance was classified into complete compliance (CC), erratic compliance (EC), and noncompliance (NC) groups. Re-examinations were carried out 11.0±0.8 years after the active periodontal treatment. The prognosis for each tooth was determined as good, questionable, or hopeless according to the bone loss observed in pretreatment radiographs. RESULTS: The rate of tooth loss of the CC group was significantly lower than that of the NC group. The tooth loss/patient and the tooth loss/patient/year were not significantly different between the three groups. The rates of tooth loss in the good, questionable, and hopeless prognosis groups were 6.7%, 9.5%, and 13.2%, respectively. For the teeth with a good prognosis, the rate of tooth loss of the CC group was significantly lower than that of the NC group (0.4% vs. 5.1%). For the teeth with a questionable prognosis, the CC group showed a significantly lower rate of tooth loss than did the EC group (4.1% vs. 30.7%) or the NC group (4.1% vs. 25.6%). For the teeth with a hopeless prognosis, the rates of tooth loss were not significantly different among the three groups. CONCLUSIONS: Within the limits of this study, the patients who showed a poor compliance with SPT were more likely to lose teeth than were the regularly compliant patients. However, the risk of tooth loss with a hopeless prognosis was high irrespective of the compliance.

A periodontitis-associated multispecies model of an oral biofilm.

PURPOSE: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. METHODS: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. RESULTS: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. CONCLUSIONS: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.

Phototoxic effect of blue light on the planktonic and biofilm state of anaerobic periodontal pathogens.

PURPOSE: The purpose of this study was to compare the phototoxic effects of blue light exposure on periodontal pathogens in both planktonic and biofilm cultures. METHODS: Strains of Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis, in planktonic or biofilm states, were exposed to visible light at wavelengths of 400.520 nm. A quartz-tungsten-halogen lamp at a power density of 500 mW/cm(2) was used for the light source. Each sample was exposed to 15, 30, 60, 90, or 120 seconds of each bacterial strain in the planktonic or biofilm state. Confocal scanning laser microscopy (CSLM) was used to observe the distribution of live/dead bacterial cells in biofilms. After light exposure, the bacterial killing rates were calculated from colony forming unit (CFU) counts. RESULTS: CLSM images that were obtained from biofilms showed a mixture of dead and live bacterial cells extending to a depth of 30-45 µm. Obvious differences in the live-to-dead bacterial cell ratio were found in P. gingivalis biofilm according to light exposure time. In the planktonic state, almost all bacteria were killed with 60 seconds of light exposure to F. nucleatum (99.1%) and with 15 seconds to P. gingivalis (100%). In the biofilm state, however, only the CFU of P. gingivalis demonstrated a decreasing tendency with increasing light exposure time, and there was a lower efficacy of phototoxicity to P. gingivalis as biofilm than in the planktonic state. CONCLUSIONS: Blue light exposure using a dental halogen curing unit is effective in reducing periodontal pathogens in the planktonic state. It is recommended that an adjunctive exogenous photosensitizer be used and that pathogens be exposed to visible light for clinical antimicrobial periodontal therapy.

Bone formation and remodeling of three different dental implant surfaces with Escherichia coli-derived recombinant human bone morphogenetic protein 2 in a rabbit model.

PURPOSE: The objective of this study was to analyze orthotropic bone formation and remodeling of three different dental implant surfaces with and without recombinant human bone morphogenetic protein 2 derived from Escherichia coli (ErhBMP-2) in a rabbit model. MATERIALS AND METHODS: Resorbable blasting media (RBM); sandblasted, large-grit, acid-etched (SLA); and magnesium-incorporated oxidized (MgO) implant surfaces were coated with ErhBMP-2 (1.5 mg/mL). The implants were placed into the proximal tibia in six New Zealand White rabbits. Each rabbit received six different implants (three coated with ErhBMP-2 in one tibia and three uncoated implants in the other tibia), and the sites were closed, submerging the implants. The animals received alizarin (at 2 weeks), calcein (at 4 weeks), and tetracycline (at 6 weeks) fluorescent bone markers, and were euthanized at 8 weeks for histomorphometric analysis. RESULTS: The amount of ErhBMP-2 coating was 9.6 ± 0.4 µg per MgO implant, 14.5 ± 0.6 µg per RBM implant, and 29.9 ± 3.8 µg per SLA implant. Clinical healing was uneventful. Mean bone-to-implant contact (± standard deviation) for the ErhBMP-2/RBM (35.4% ± 5.1%) and ErhBMP-2/MgO (33.4 % ± 13.2%) implants was significantly greater compared with RBM (23.6% ± 6.2%) and MgO (24.9% ± 2.7%) implants (P < .05). Considering the mean bone-to-implant contact in cortical bone, ErhBMP-2/SLA implants (32.9% ± 7.8%) showed lower bone-to-implant contact in cortical bone than all other implant variations (range, 39.9% ± 18.1% to 51.3% ± 9.2%; P < .05). There were no remarkable differences in new bone area, with minor differences between implants. CONCLUSIONS: Within the limits of study, it was found that the absorbed ErhBMP-2 dose varied with implant surface characteristics, influencing local bone formation and remodeling.

Masticatory function following implants replacing a second molar.

PURPOSE: The aim of this study was to obtain objective and standardized information on masticatory function and patient satisfaction following second molar single implant therapy. METHODS: Twenty adult patients, who had restored second molar single implants more than 1 month before the study, were enrolled in this study. All patients received a chewing test using peanuts before and after insertion of the implant prosthesis, with a questionnaire and visual analogue scale (VAS) to evaluate the effect of second molar single implant therapy. RESULTS: This study obtained standardized information on the masticatory function objectively (e.g., P, R, X(50)) before (Pre-insertion) and after insertion (Post-insertion) of the implant prosthesis. Masticatory performance (P) after insertion of the implant prosthesis significantly increased from 67.8±9.9 to 84.3±8.5% (P<0.0001). With the implant prosthesis, the P value increased by 24%. The masticatory efficiency index (R) of Post-insertion is higher than that of Pre-insertion (P<0.0001). With the implant prosthesis, the R value increased by 29%. The median particle size (X(50)) of Post-insertion is lower than that of Pre-insertion (P<0.0001). More than 90% of the patients were satisfied with the second molar single implant therapy from a functional point of view. CONCLUSIONS: These findings indicate that a second molar single implant can increase masticatory function.