Implant stability at the time of surgery is crucial for the long-term success of dental implants. Primary stability is considered of paramount importance to achieve osseointegration. The purpose of the present study was to investigate the correlation between the insertion torque and primary stability of dental implants using artificial bone blocks with different bone densities and compositions to mimic different circumstances that are encountered in routine daily clinical settings.
In order to validate the objectives, various sized holes were made in bone blocks with different bone densities (#10, #20, #30, #40, and #50) using a surgical drill and insertion torque together with implant stability quotient (ISQ) values that were measured using the Osstell Mentor. The experimental groups under evaluation were subdivided into 5 subgroups according to the circumstances.
In group 1, the mean insertion torque and ISQ values increased as the density of the bone blocks increased. For group 2, the mean insertion torque values decreased as the final drill size expanded, but this was not the case for the ISQ values. The mean insertion torque values in group 3 increased with the thickness of the cortical bone, and the same was true for the ISQ values. For group 4, the mean insertion torque values increased as the cancellous bone density increased, but the correlation with the ISQ values was weak. Finally, in group 5, the mean insertion torque decreased as the final drill size increased, but the correlation with the ISQ value was weak.
Within the limitations of the study, it was concluded that primary stability does not simply depend on the insertion torque, but also on the bone quality.
Bone density; Dental implants; Torque
The characteristics of the T cell response to the members of oral flora are poorly understood. We characterized the antibody and T cell responses to FadA and Td92, adhesins from Fusobacterium nucleatum, an oral commensal, and Treponema denticola, a periodontal pathogen, respectively. Peripheral blood and saliva were obtained from healthy individuals and patients with untreated chronic periodontitis (CP, n = 11 paris) and after successful treatment of the disease (n = 9). The levels of antigen-specific antibody were measured by ELISA. In plasma, IgG1 was the most abundant isotype of Ab for both Ags, followed by IgA and then IgG4. The levels of FadA-specific salivary IgA (sIgA) were higher than Td92-specific sIgA and the FadA-specific IgA levels observed in plasma. However, the periodontal health status of the individuals did not affect the levels of FadA- or Td92-specific antibody. Even healthy individuals contained FadA- and Td92-specific CD4+ T cells, as determined by the detection of intracytoplasmic CD154 after short-term in vitro stimulation of peripheral blood mononuclear cells (PBMCs) with the antigens. Patients with CP tended to possess increased numbers of FadA- and Td92-specific CD4+ T cells but reduced numbers of Td92-specific Foxp3+CD4+ Tregs than the healthy subjects. Both FadA and Td92 induced the production of IFNγ and IL-10 but inhibited the secretion of IL-4 by PBMCs. In conclusion, F. nucleatum induced Th3 (sIgA)- and Th1 (IFNγ and IgG1)-dominant immune responses, whereas T. denticola induced a Th1 (IFNγ and IgG1)-dominant response. This IFNγ-dominant cytokine response was impaired in CP patients, and the Td92-induced IFNγ levels were negatively associated with periodontal destruction in patients. These findings may provide new insights into the homeostatic interaction between the immune system and oral bacteria and the pathogenesis of periodontitis.
The aim of this study was to investigate the effects of synthetic fibronectin (FN) fragments, including fibrin binding sites from amino-terminal FN fragments containing type I repeats 1 to 5, on osteoblast-like cell activity.
Oligopeptides ranging from 9 to 20 amino acids, designated FF1, FF3, and FF5, were synthesized by a solid-phase peptide synthesizing system, and we investigated the effects of these peptides on cell attachment and extent of mineralization using confocal microscopy, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, and Alizarin red S staining.
FF3 and FF5 peptides increased the number of attached human osteoblastic cells, and FF3 administration led to prominent cell spreading. Mineralization was increased in FF3 and FF5 compared to FF1 and the untreated control.
Taken together, it can be concluded that the fibrin-binding oligopeptides FF3 and FF5 enhanced cell attachment and mineralization on osteoblast-like cells. These results indicate that FF3 and FF5 have the potential to increase osteoblast-like cell activity. Performing an in vivo study may provide further possibilities for surface modification of biomimetic peptides to enhance osteogenesis, thus improving the regeneration of destroyed alveolar bone.
Fibrin; Fibronectins; Oligopeptides; Osteoblasts
The purpose of this study was to investigate the effects of the immunosuppressants FK506 and cyclosporin A (CsA) on the osteogenic differentiation of rat mesenchymal stem cells (MSCs).
The effect of FK506 and CsA on rat MSCs was assessed in vitro. The MTT assay was used to determine the deleterious effect of immunosuppressants on stem cell proliferation at 1, 3, and 7 days. Alkaline phosphatase (ALP) activity was analyzed on days 3, 7, and 14. Alizarin red S staining was done on day 21 to check mineralization nodule formation. Real-time polymerase chain reaction (RT-PCR) was also performed to detect the expressions of bone tissue-specific genes on days 1 and 7.
Cell proliferation was promoted more in the FK506 groups than the control or CsA groups on days 3 and 7. The FK506 groups showed increased ALP activity compared to the other groups during the experimental period. The ALP activity of the CsA groups did not differ from the control group in any of the assessments. Mineralization nodule formation was most prominent in the FK506 groups at 21 days. RT-PCR results of the FK506 groups showed that several bone-related genes-osteopontin, osteonectin, and type I collagen (Col-I)-were expressed more than the control in the beginning, but the intensity of expression decreased over time. Runx2 and Dlx5 gene expression were up-regulated on day 7. The effects of 50 nM CsA on osteonectin and Col-I were similar to those of the FK506 groups, but in the 500 nM CsA group, most of the genes were less expressed compared to the control.
These results suggest that FK506 enhances the osteoblastic differentiation of rat MSCs. Therefore, FK506 might have a beneficial effect on bone regeneration when immunosuppressants are needed in xenogenic or allogenic stem cell transplantation to treat bone defects.
Cell differentiation; Cyclosporin A; FK506; Immunosuppressive agents; Mesenchymal stem cells
The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2.
MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days.
At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7.
The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.
Bone morphogenetic protein-2; Cell differentiation; Cell proliferation; Zirconium oxide
The aim of this study is to determine whether certain biomaterials have the potential to support cell attachment. After seeding bone marrow stromal cells onto the biomaterials, we investigated their responses to each material in vitro.
Rat bone marrow derived stromal cells were used. The biomaterials were deproteinized bovine bone mineral (DBBM), DBBM coated with fibronectin (FN), synthetic hydroxyapatite (HA), HA coated with FN, HA coated with β-tricalcium phosphate (TCP), and pure β-TCP. With confocal laser scanning microscopy, actin filaments and vinculin were observed after 6, 12, and 24 hours of cell seeding. The morphological features of cells on each biomaterial were observed using scanning electron microscopy at day 1 and 7.
The cells on HA/FN and HA spread widely and showed better defined actin cytoskeletons than those on the other biomaterials. At the initial phase, FN seemed to have a favorable effect on cell adhesion. In DBBM, very few cells adhered to the surface.
Within the limitations of this study, we can conclude that in contrast with DBBM not supporting cell attachment, HA provided a more favorable environment with respect to cell attachment.
Bone substitutes; Cell adhesion; Fibronectins; Stem cells
The aim of this study was to investigate the immunomodulatory effects of canine periodontal ligament stem cells on allogenic and xenogenic immune cells in vitro.
Mixed cell cultures consisting of canine stem cells (periodontal ligament stem cells and bone marrow stem cells) and allogenic canine/xenogenic human peripheral blood mononuclear cells (PBMCs) were established following the addition of phytohemagglutinin. The proliferation of PBMCs was evaluated using the MTS assay. The cell division of PBMCs was analyzed using the CFSE assay. The apoptosis of PBMCs was assessed using the trypan blue uptake method.
Periodontal ligament stem cells and bone marrow stem cells inhibited the proliferation of allogenic and xenogenic PBMCs. Both periodontal ligament stem cells and bone marrow stem cells suppressed the cell division of PBMCs despite the existence of a mitogen. No significant differences in the percentages of apoptotic PBMCs were found among the groups.
Canine periodontal ligament stem cells have an immunomodulatory effect on allogenic and xenogenic PBMCs. This effect is not a product of apoptosis of PBMCs but is caused by the inhibition of cell division of PBMCs.
Allogenic; Immunomodulation; Periodontal ligaments; Stem cell; Xenogenic
Fibronectin (FN) has been shown to stimulate bone regeneration in animal models. The aim of this study was to evaluate the capacity of bovine bone mineral coated with synthetic oligopeptides to enhance bone regeneration in rabbit calvarial defects.
Oligopeptides including fibrin-binding sequences of FN repeats were synthesized on the basis of primary and tertiary human plasma FN structures. Peptide coated and uncoated bone minerals were implanted into 10 mm calvarial defects in New Zealand white rabbits, and the animals were sacrificed at 4 or 8 weeks after surgery. After specimens were prepared, histologic examination and histomorphometric analysis were performed.
At 4 weeks after surgery, the uncoated groups showed a limited amount of osteoid formation at the periphery of the defect and the oligopeptide coated groups showed more osteoid formation and new bone formation in the center of the defect as well as at the periphery. At 8 weeks, both sites showed increased new bone formation. However, the difference between the two sites had reduced.
Fibrin-binding synthetic oligopeptide derived from FN on deproteinized bovine bone enhanced new bone formation in rabbit calvarial defects at the early healing stage. This result suggests that these oligopeptides can be beneficial in reconstructing oral and maxillofacial deformities or in regenerating osseous bone defects.
Fibrin; Fibronectins; Oligopeptides
The healing process following tooth extraction apparently results in a pronounced resorption of the alveolar ridge. As a result, the width of alveolar ridge is reduced and severe alveolar bone resorption occurs. The purpose of this experiment is to clinically and histologically evaluate the results of using horse-derived bone mineral for socket preservation.
The study comprised 4 patients who were scheduled for extraction as a consequence of severe chronic periodontitis or apical lesion. The extraction was followed by socket preservation using horse-derived bone minerals. Clinical parameters included buccal-palatal width, mid-buccal crest height, and mid-palatal crest height. A histologic examination was conducted.
The surgical sites healed uneventfully. The mean ridge width was 7.75 ± 2.75 mm at baseline and 7.00 ± 2.45 mm at 6 months. The ridge width exhibited no significant difference between baseline and 6 months. The mean buccal crest height at baseline was 7.5 ± 5.20 mm, and at 6 months, 3.50 ± 0.58 mm. The mean palatal crest height at baseline was 7.75 ± 3.10 mm, and at 6 months, 5.00 ± 0.82 mm. There were no significant differences between baseline and 6 months regarding buccal and palatal crest heights. The amount of newly formed bone was 9.88 ± 2.90%, the amount of graft particles was 42.62 ± 6.57%, and the amount of soft tissue was 47.50 ± 9.28%.
Socket preservation using horse-derived bone mineral can effectively maintain ridge dimensions following tooth extraction and can promote new bone formation through osteoconductive activities.
Bone resorption; Bone substitutes; Clinical trial; Tooth socket
To prolong the degradation time of collagen membranes, various cross-linking techniques have been developed. For cross-linking, chemicals such as formaldehyde and glutaraldehyde are added to collagen membranes, but these chemicals could adversely affect surrounding tissues. The aim of this study is to evaluate the ability of porous non-chemical cross-linking porcine-derived collagen nanofibrous membrane to enhance bone and associated tissue regeneration in one-wall intrabony defects in beagle dogs.
The second and third mandibular premolars and the first molars of 2 adult beagles were extracted bilaterally and the extraction sites were allowed to heal for 10 weeks. One-wall intrabony defects were prepared bilaterally on the mesial and distal side of the fourth mandibular premolars. Among eight defects, four defects were not covered with membrane as controls and the other four defects were covered with membrane as the experimental group. The animals were sacrificed 10 weeks after surgery.
Wound healing was generally uneventful. For all parameters evaluating bone regeneration, the experimental group showed significantly superior results compared to the control. In new bone height (NBh), the experimental group exhibited a greater mean value than the control (3.04 ± 0.23 mm/1.57 ± 0.59, P = 0.003). Also, in new bone area (NBa) and new bone volume (NBv), the experimental group showed superior results compared to the control (NBa, 34.48 ± 10.21% vs. 5.09 ± 5.76%, P = 0.014; and NBv, 28.04 ± 12.96 vs. 1.55 ± 0.57, P = 0.041). On the other hand, for parameters evaluating periodontal tissue regeneration, including junctional epithelium migration and new cementum height, there were no statistically significant differences between two groups.
Within the limitations of this study, this collagen membrane enhanced bone regeneration at one-wall intrabony defects. On the other hand, no influence of this membrane on periodontal tissue regeneration could be ascertained in this study.
Absorbable implants; Bone regeneration; Collagen; Guided tissue regeneration
It has been reported that low-level semiconductor diode lasers could enhance the wound healing process. The periodontal ligament is crucial for maintaining the tooth and surrounding tissues in periodontal wound healing. While low-level semiconductor diode lasers have been used in low-level laser therapy, there have been few reports on their effects on periodontal ligament fibroblasts (PDLFs). We performed this study to investigate the biological effects of semiconductor diode lasers on human PDLFs.
Human PDLFs were cultured and irradiated with a gallium-aluminum-arsenate (GaAlAs) semiconductor diode laser of which the wavelength was 810 nm. The power output was fixed at 500 mW in the continuous wave mode with various energy fluencies, which were 1.97, 3.94, and 5.91 J/cm2. A culture of PDLFs without laser irradiation was regarded as a control. Then, cells were additionally incubated in 72 hours for MTS assay and an alkaline phosphatase (ALPase) activity test. At 48 hours post-laser irradiation, western blot analysis was performed to determine extracellular signal-regulated kinase (ERK) activity. ANOVA was used to assess the significance level of the differences among groups (P<0.05).
At all energy fluencies of laser irradiation, PDLFs proliferation gradually increased for 72 hours without any significant differences compared with the control over the entire period taken together. However, an increment of cell proliferation significantly greater than in the control occurred between 24 and 48 hours at laser irradiation settings of 1.97 and 3.94 J/cm2 (P<0.05). The highest ALPase activity was found at 48 and 72 hours post-laser irradiation with 3.94 J/cm2 energy fluency (P<0.05). The phosphorylated ERK level was more prominent at 3.94 J/cm2 energy fluency than in the control.
The present study demonstrated that the GaAlAs semiconductor diode laser promoted proliferation and differentiation of human PDLFs.
Alkaline phosphatase; Extracellular signal-regulated kinases; Fibroblasts; Periodontal ligament; Semiconductor diode lasers
The purpose of this study was to compare the bone regeneration effects of cortical, cancellous, and cortico-cancellous human bone substitutes on calvarial defects of rabbits.
Four 8-mm diameter calvarial defects were created in each of nine New Zealand white rabbits. Freeze-dried cortical bone, freeze-dried cortico-cancellous bone, and demineralized bone matrix with freeze-dried cancellous bone were inserted into the defects, while the non-grafted defect was regarded as the control. After 4, 8, and 12 weeks of healing, the experimental animals were euthanized for specimen preparation. Micro-computed tomography (micro-CT) was performed to calculate the percent bone volume. After histological evaluation, histomorphometric analysis was performed to quantify new bone formation.
In micro-CT evaluation, freeze-dried cortico-cancellous human bone showed the highest percent bone volume value among the experimental groups at week 4. At week 8 and week 12, freeze-dried cortical human bone showed the highest percent bone volume value among the experimental groups. In histologic evaluation, at week 4, freeze-dried cortico-cancellous human bone showed more prominent osteoid tissue than any other group. New bone formation was increased in all of the experimental groups at week 8 and 12. Histomorphometric data showed that freeze-dried cortico-cancellous human bone showed a significantly higher new bone formation percentile value than any other experimental group at week 4. At week 8, freeze-dried cortical human bone showed the highest value, of which a significant difference existed between freeze-dried cortical human bone and demineralized bone matrix with freeze-dried cancellous human bone. At week 12, there were no significant differences among the experimental groups.
Freeze-dried cortico-cancellous human bone showed swift new bone formation at the 4-week healing phase, whereas there was less difference in new bone formation among the experimental groups in the following healing phases.
Bone Substitutes; Osteogenesis; Transplantation; X-Ray Microtomography
It has been suggested that primary implant stability plays an essential role in successful osseointegration. Resonance frequency analysis (RFA) is widely used to measure the initial stability of implants because it provides superior reproducibility and non-invasiveness. The purpose of this study is to investigate whether the fractal dimension from the panoramic radiograph is related to the primary stability of the implant as represented by RFA.
This study included 22 patients who underwent dental implant installation at the Department of Periodontology of Seoul National University Dental Hospital. Morphometric analysis and fractal analysis of the bone trabecular pattern were performed using panoramic radiographs, and the implant stability quotient (ISQ) values were measured after implant installation using RFA. The radiographs of 52 implant sites were analyzed, and the ISQ values were compared with the results from the morphometric analysis and fractal analysis.
The Pearson correlation showed a linear correlation between the ISQ values of RFA and the parameters of morphometric analysis but not of statistical significance. The fractal dimension had a linear correlation that was statistically significant. The correlation was more pronounced in the mandible.
In conclusion, we suggest that the fractal dimension acquired from the panoramic radiograph may be a useful predictor of the initial stability of dental implants.
Dental implantation; Fractals; Panoramic radiography