The local administration of parathyroid hormone encourages the healing of bone defects in the rat calvaria: Micro-computed tomography,histological and histomorphometric evaluation

ObjectiveTo evaluate the effect of a single-dose local administration of PTH on bone healing in rat calvarial bone defects by means of micro-computed tomography, histological and histomorphometric analysis.DesignCritical-size cranial osteotomy defects were created in 42 male rats. The animals were randomly divided into 3 groups. In the C Group, the bone defect was only filled with a blood clot. In the S Group, it was filled with a collagen sponge and covered with bovine cortical membrane. In the PTH Group, the defect was filled with a collagen sponge soaked with PTH and covered with bovine cortical membrane. The groups were further split in two for euthanasia 15 and 60 days post-surgery. Data was statistically analyzed with t-tests for independent samples or the nonparametric Mann-Whitney test when applicable. Intragroup comparisons were analyzed with paired t-tests (p < 0.05).ResultsMicro-CT analysis results did not demonstrate statistically significant intergroup differences. At 15 days post-surgery, the histomorphometric analysis showed that the PTH Group exhibited a significantly higher percentage of bone formation compared with the S Group. At 60 days post-surgery, a higher percentage of new bone was observed in the PTH group.ConclusionThe results suggest that the local administration of PTH encouraged the bone healing in critical-size calvarial defects in rats.     

Inhibitory effect of 1,25-dihydroxyvitamin D3 on Porphyromonas gingivalis-induced inflammation and bone resorption in vivo

ObjectiveTo investigate whether intragastric administration of 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) could inhibit the bone resorption and inflammation in a mouse calvarial model infected by Porphyromonas gingivalis (P. gingivalis).DesignLive P. gingivalis ATCC 33277 was injected once daily for 6 days into the subcutaneous tissue overlying the calvaria in mice. At the same time, 1,25(OH)₂D₃ (50 μg/kg per day) was administered by gavage for 9 days, starting 3 d before the infection. Mice were killed under ether anesthesia 8 h after the last injection of P. gingivalis. Micro-computed tomography scanning was used to evaluate calvarial bone loss. Tartrate-resistant acid phosphatase was used to detect osteoclast activity. Real-time PCR was used to assess the mRNA expressions of OPG, RANKL, c-Fos, NFATc1, CTSK and TRAP in calvarial bone and IL-6, IL-10, IL-1β, IL-12p40 and TNF-α in soft tissue. The levels of serum IL-6, IL-10 were determined by ELISA.Results1,25(OH)₂D₃ treatment apparently attenuated bone resorption in P. gingivalis-induced mouse calvarial model and markedly reduced the number of osteoclasts. The expression levels of RANKL and osteoclast-related genes such as c-Fos, NFATc1, CTSK and TRAP were also decreased by 1,25(OH)₂D₃. Besides, 1,25(OH)₂D₃ inhibited the expression of pro-inflammatory cytokines IL-6, IL-12p40 and TNF-α and enormously elevated the expression of anti-inflammatory cytokine IL-10.Conclusion1,25(OH)₂D₃ may decrease bone resorption in vivo via suppressing the expression of osteoclast-related genes and its anti-inflammatory properties.     

Development and evaluation of novel biodegradable chitosan based metformin intrapocket dental film for the management of periodontitis and alveolar bone loss in a rat model

ObjectiveThe aim of this study was to develop a chitosan-metformin based intrapocket dental film (CMIDF) for applications in the treatment of periodontitis and alveolar bone loss in an rat model of periodontitis.DesignCMIDF inserts were fabricated by the solvent casting technique. The fabricated inserts were evaluated for physical characteristics such as folding endurance, surface pH, mucoadhesive strength, metformin content uniformity, and release. X-ray diffraction analysis indicates no crystallinity of metformin in presence of chitosan which confirmed successful entrapment of metformin into the CMIDF. Fourier-transform infrared spectroscopy revealed stability of CMIDF and compatibility between metformin and chitosan. Periodontitis was induced by a combination of Porphyromonas gingivalis- lipopolysaccharide injections in combinations with ligatures around the mandibular first molar. We divided rats into 5 groups (8 rats/group): healthy, untreated periodontitis; periodontitis plus CMIDF-A (1.99 ± 0.09 mg metformin; total mass-4.01 ± 0.05 mg), periodontitis plus CMIDF-B (2.07 ± 0.06 mg metformin; total mass-7.56 ± 0.09 mg), and periodontitis plus chitosan film (7.61 ± 0.08 mg). After four weeks, mandibles were extracted to evaluate alveolar bone loss by micro-computerized tomography and histological techniques.ResultsAlveolar bone was intact in the healthy group. Local administration of CMIDF resulted in significant improvements in the alveolar bone properties when compared to the untreated periodontitis group. The study reported here demonstrates that novel CMIDF showed good antibacterial activity and effectively reduced alveolar bone destruction in a rat model of experimental periodontitis.ConclusionsNovel CMIDF showed good antibacterial activity and improved alveolar bone properties in a rat model.     

Induction of immune response and prevention of alveolar bone loss with recombinant Porphyromonas gingivalis peptidylarginine deiminase

ObjectiveTreatments for periodontitis are not absolutely perfect, and a vaccine against Porphyromonas gingivalis (P. gingivalis) could become a valuable adjunct therapy for periodontitis.DesignIn this study, a vaccine of peptidylarginine deiminase (PAD) from P. gingivalis was evaluated in P. gingivalis-induced murine lesion and periodontitis models. The prevention of alveolar bone loss analysis determined by micro-computed X-ray tomography (micro-CT), and histological assays. Furthermore, the induction of immune response of mouse anti-PAD done with ELISA and Western Blot analysis.ResultsCompared with animal immunization with incomplete Freund's adjuvant (IFA) alone, PAD group significantly inhibited (P < 0.05) bone resorption. ELISA and Western Blot showed that PAD induced response involving immunoglobulin G1 (Ig G1) predominantly.ConclusionsThese results suggest that PAD could be a candidate antigen for a vaccine against P. gingivalis infection.     

Adhesion of Porphyromonas gingivalis and Tannerella forsythia to dentin and titanium with sandblasted and acid etched surface coated with serum and serum proteins – An in vitro study

ObjectiveTo evaluate the adhesion of selected bacterial strains incl. expression of important virulence factors at dentin and titanium SLA surfaces coated with layers of serum proteins.MethodsDentin- and moderately rough SLA titanium-discs were coated overnight with human serum, or IgG, or human serum albumin (HSA). Thereafter, Porphyromonas gingivalis, Tannerella forsythia, or a six-species mixture were added for 4 h and 24 h. The number of adhered bacteria (colony forming units; CFU) was determined. Arg-gingipain activity of P. gingivalis and mRNA expressions of P. gingivalis and T. forsythia proteases and T. forsythia protease inhibitor were measured.ResultsCoating specimens never resulted in differences exceeding 1.1 log10 CFU, comparing to controls, irrespective the substrate. Counts of T. forsythia were statistically significantly higher at titanium than dentin, the difference was up to 3.7 log10 CFU after 24 h (p = 0.002). No statistically significant variation regarding adhesion of the mixed culture was detected between surfaces or among coatings. Arg-gingipain activity of P. gingivalis was associated with log10 CFU but not with the surface or the coating. Titanium negatively influenced mRNA expression of T. forsythia protease inhibitor at 24 h (p = 0.026 uncoated, p = 0.009 with serum).ConclusionsThe present findings indicate that: a) single bacterial species (T. forsythia) can adhere more readily to titanium SLA than to dentin, b) low expression of T. forsythia protease inhibitor may influence the virulence of the species on titanium SLA surfaces in comparison with teeth, and c) surface properties (e.g. material and/or protein layers) do not appear to significantly influence multi-species adhesion.     

Frequency of periodontal pathogens in equivalent peri-implant and periodontal clinical statuses

ObjectivesThis study tested the hypotheses that there is: (1) higher bacterial frequency in peri-implantitis/periodontitis, followed by mucositis/gingivitis and peri-implant/periodontal health; (2) similar bacterial frequency between comparable peri-implant and periodontal clinical statuses.Design of studyThe presence of Porphyromonas gingivalis, Tannerella forsythia, Campylobacter rectus, Prevotella intermedia, Treponema denticola and Aggregatibacter actinomycetemcomitans was evaluated in peri-implant (n = 53) and periodontal (n = 53) health; mucositis (n = 50), gingivitis (n = 50), peri-implantitis (n = 50) and periodontitis (n = 50).ResultsThe pattern of peri-implant bacterial frequency was not as expected (peri-implantitis > mucositis > health). Except for P. intermedia (p > 0.05), bacterial frequency was higher in peri-implantitis than health (p < 0.05). The frequency of P.gingivalis and red complex species were higher in peri-implantitis than mucositis (p < 0.05). In periodontal samples, T. forsythia and T. denticola showed the expected pattern of frequency (periodontitis > gingivitis > health). The frequencies of C. rectus and T. forsythia were higher in healthy teeth/gingivitis than healthy implants/mucositis, respectively (p < 0.05). The frequency of P. gingivalis and A. actinomycetemcomitans were similar between periodontitis and peri-implantitis (p > 0.05) while all other species occurrences were higher in periodontitis than peri-implantitis (p < 0.05).ConclusionsBacterial frequency increased from peri-implant/periodontal health to peri-implantitis/periodontitis but not from mucositis/gingivitis to peri-implantitis/periodontitis. There was a trend towards higher bacterial frequency in teeth than implants.     

Effects of connective tissue growth factor on human periodontal ligament fibroblasts

ObjectiveThe aim of this study was to evaluate the effects of different concentrations of connective tissue growth factor (CTGF) on human periodontal ligament fibroblasts(HPLFs).DesignHPLFs were cultured and identified. Then, different concentrations of CTGF (1, 5, 10, 50, 100 ng/ml) were added to the HPLF culture. Next, CCK-8 assays, alkaline phosphatase (ALP) assays, hydroxyproline determination, alizarin red staining methods, Transwell chambers and real-time PCR methods were applied to observe the effects of CTGF on the proliferation, ALP activity, synthesis of collagen, formation of mineralized nodules and migration. We also studied expression of ALP, fiber link protein (FN), integrin-binding sialoprotein (IBSP), osteocalcin (OC), and integrin beta 1 (ITGB1) mRNA by HPLFs. Statistical significance was assumed if P < 0.05 or P < 0.01.ResultsThe addition of CTGF (1, 5, 10 ng/ml) remarkably promoted the proliferation and collagen synthesis of HPLFs compared with controls. CTGF (1, 5, 10, 50 ng/ml) improved ALP activity of HPLFs, and at all concentrations, CTGF (1, 5, 10, 50, 100 ng/ml) improved the expression of ALP, FN, IBSP and ITGB1 mRNA. In addition, CTGF (1, 5, 10, 50, 100 ng/ml) promoted the migration of HPLFs, which was dose-dependent, with maximal promotion in the 10 ng/ml group (P < 0.05 or P < 0.01).ConclusionsThus, in a certain range of concentrations, CTGF can promote the biological effects, including proliferation, migration and collagen synthesis of HPLFs, to promote the differentiation of HPLFs in the process of osteogenesis.     

Maxillary sinus floor augmentation and dental implant placement using dentin matrix protein-1 gene-modified bone marrow stromal cells mixed with deproteinized boving bone: A comparative study in beagles

ObjectiveThe aim of the study was to evaluate the effects of the combined use of dentin matrix protein-1 (DMP1) gene-modified bone marrow stromal cells (BMSCs) and Bio-Oss^® for maxillary sinus floor augmentation (MSFA) implant placement in dogs.Materials and methodsBMSCs were derived from bone marrow of six beagles and cultured. The cells were transduced with a lentiviral vector overexpressing the DMP1 gene and enhanced green fluorescent protein (EGFP) gene (Lenti-DMP1/EGFP) in test group, and with a lentiviral vector encoding EGFP gene (Lenti-EGFP) in control group. Six dogs received sinus augmentations using the bilateral approach with a simultaneous implant placement at each site respectively. At the same concentration, 2 × 10⁷ cells/ml, one sinus was grafted using a mixture of autologous DMP1/EGFP gene-modified BMSCs and Bio-Oss^® (DMP1 group), and the contralateral sinus was grafted with autologous EGFP gene-modified bMSCs and Bio-Oss^® (EGFP group). After a 3 month healing period, bone regeneration and osseointegration were evaluated using histologic and histomorphometric methods.ResultsThe bone-implant contact (BIC) and the bone area fraction in the DMP1 group (BIC: 34.67% ± 8.23%, bone area fraction: 35.16% ± 3.32%) were significantly greater compared with the EGFP group (BIC: 26.06% ± 5.16%, bone area fraction: 20.74% ± 1.63%) (P < 0.05). No significant difference between the residual bone substitute material volume (BSMV) in the DMP1 group (35.86 ± 7.35) and the EGFP group (32.16 ± 9.16) was found in our study (P > 0.05).ConclusionBMSCs modified with the DMP1 gene can be used as an adjunct to Bio-Oss^® to enhance new bone formation and the osseointegration of dental implants in MSFA of dogs.     

A collagen membrane containing osteogenic protein-1 facilitates bone regeneration in a rat mandibular bone defect

ObjectivesOsteogenic protein-1 (OP-1) has shown osteoinductive activities and is useful for clinical treatments, including bone regeneration. Regenerative procedures using a bioabsorbable collagen membrane (BCM) are well established in periodontal and implant dentistry. We evaluated the subsequent effects of the BCM in combination with OP-1 on bone regeneration in a rat mandibular circular critical-sized bone defect in vivo.DesignWe used 8 rats that received surgery in both sides of the mandible, and created the total 16 defects which were divided into 4 groups: Group 1; no treatment, as a control, Group 2; BCM alone, Group 3; BCM containing low dose 0.5 μg of OP-1 (L-OP-1), and Group 4; BCM containing high dose 2.0 μg of OP-1 (H-OP-1). Newly formed bone was evaluated by micro computed tomography (micro-CT) and histological analyses at 8 weeks postoperatively. In quantitative and qualitative micro-CT analyses of the volume of new bone formation, bone density, and percentage of new bone area was evaluated.ResultsBCM with rhOP-1 significantly increased and accelerated bone volume, bone mineral density, and percentage of new bone area compared to control and BCM alone at 8 weeks after surgery; these enhancements in bone regeneration in the OP-1-treated groups were dose-dependent.ConclusionsOP-1 delivered with a BCM may have effective osteoinductive potency and be a good combination for bone regeneration. The use of such a combination device for osteogenesis may result in safer and more predictable bone regenerative outcomes in the future.     

The effect of residual bone height and membrane thickness on sinus membrane perforation in crestal sinus grafting: A prospective clinical study

This study sought to determine the rate of sinus membrane perforation in patients undergoing crestal sinus grafting, as well as the effect of Schneiderian membrane thickness and residual bone height (RBH) on membrane perforation, using cone beam computed tomography. The study included 25 patients undergoing 44 crestal sinus grafting procedures. The sites for crestal sinus grafting were divided into a control group (RBH ≥ 5 mm) and a test group (RBH < 5 mm). All sinus grafting procedures were also categorised based on membrane thickness: group A (<1 mm), group B (1–2 mm), and group C (≥2 mm). The rate of membrane perforation was 18.2%. The median RBH measurement was 5.59 mm. No statistically significant difference in membrane perforation rate was found between the test and control groups (P = 0.262). The median thickness of the Schneiderian membrane was 1.35 mm. There was no statistically significant difference in membrane perforation among the three membrane thickness groups (P = 0.431). No significant correlation between RBH and membrane perforation was observed, although clinical observation indicated that there was a tendency for an increased membrane perforation rate in the presence of a RBH < 5 mm. The perforation rate was found to be at its highest when the membrane was thinner than 1 mm.     

Effect of proanthocyanidin on ultrastructure and mineralization of dentine collagen

ObjectiveProanthocyanidin (PA) is a natural collagen cross-linker that has been used in dentine matrix biomodification for reparative and preventive therapies. This study evaluated the ultrastructure of collagen after its interaction with PA. Furthermore, the mineralization of PA-biomodified collagen matrix was observed.MethodsTen freshly extracted sound human molars were sectioned into 0.5 mm × 1.7 mm × 7 mm beams for ultrastructural evaluation of PA and dentine matrix under Field Emission Scanning Electron Microscopy (FESEM) and Transmission Electron Microscopy (TEM). Specimens for TEM were completely demineralized and divided into three groups according to PA treatments: deionized water, 2% PA and 6.5% PA. The specimens were fixed, dehydrated, sectioned and examined using TEM. Specimens for FESEM were lightly conditioned with EDTA and similarly divided into the three groups for observation using FESEM. Type I collagen from calf skin was used to analyse the mineral interaction after treatment with 6.5% PA. Formvar- and carbon-coated 400-mesh Ni grids (EMS, Hatfiels, PA, USA) were placed over a 2 mg/mL collagen solution prepared from calf skin-derived Type I collagen to achieve self-assembly of collagen fibrils. Grids were treated with 6.5% PA and divided into two groups. One group was floated over a remineralization solution containing 20 mM HEPES, 2.25 mM CaCl₂-2H₂O, 1.35 mM KH₂PO4, 3.08 mM NaN3 and 130 mM KCl and the other group was over a CPP-ACP solution (Tooth mousse 1:100 dilution with deionized water). The floating samples were kept in a 37 °C and 100% humidity chamber. Grids were taken out at selected time durations (24 h, 48 h and 72 h for mineralization solution/24 h for CPP-ACP) and observed under TEM without staining. Selected area electron diffractions (SAEDs) were performed at 110 kV.ResultsFollowing treatment of demineralized dentine collagen matrix with PA, the size and number of interfibrillar spaces were reduced. The collagen fibrils aggregated together with a reduction in porosity. A characteristic banding pattern of collagen fibrils was observed under TEM. Treatment of PA-biomodified collagen fibrils with remineralization solution increased mineral aggregation along its long axis, when compared to the control group. Furthermore, treatment of PA-biomodified collagen fibrils with CPP-ACP solution enhanced mineral uptake and deposition as well as initiated apatite formation within 24 h.ConclusionProanthocyanidin alters the ultrastructure of demineralized dentine collagen matrix. The PA-biomodified collagen matrix promotes remineralization.     

The effect of nicotine on osteoinduction by recombinant human bone morphogenetic protein 2

Nicotine, one of the constituents of tobacco, is known to have an adverse effect on human health. We sought to clarify the interaction between nicotine and recombinant human bone morphogenetic protein 2 (rhBMP-2) in terms of osteogenesis in vitro and osteoinduction in vivo. Nicotine did not inhibit or stimulate alkaline phosphatase (ALP) activity or the amount of osteocalcin in C2C12 cells in the presence of rhBMP-2 in vitro. Ectopic bone formation using a collagen sponge containing rhBMP-2 was evaluated with and without nicotine after 21 days using radiographic, histological, biochemical, and immunohistochemical analyses. ALP activity in the medium-dose group (2.2 ± 0.9 IU/mg protein; P = 0.047) and the high-dose group (2.0 ± 0.1 IU/mg protein; P = 0.03) was significantly lower than in the control group. The calcium content in the medium-dose group (35.4 ± 12.9 μg/mg tissue; P = 0.0099) and high-dose group (34.8 ± 10.5 μg/mg tissue; P = 0.006) was significantly lower than in the control group. The number of vascular endothelial growth factor-positive cells in the high-dose group (671.9 ± 57.3 cells/mm²; P = 0.03) was significantly lower than in the control group. Results showed that nicotine did not inhibit the stimulatory effect of rhBMP-2 in vitro, but a high dose of nicotine inhibited bone formation in vivo by adversely affecting vascularization.     

In vitro failure and fracture resistance of veneered and full-contour zirconia restorations

ObjectivesThis study evaluated the failure and fracture resistance of zirconia-based fixed partial dentures (FPDs) under the influence of different surface treatments and adjustment procedures.MethodsSeven groups (n = 8/group) of three-unit zirconia-based FPDs were fabricated in anatomic design (AD) or anatomically reduced design (ARD) and surfaces were prepared according to clinical relevance: #1: AD – sintered; #2: AD – sintered – glazed; #3: AD – sintered – sandblasted – glazed; #4: AD – sintered – polished – grinded (contact points adjusted); #5: AD – sintered – polished – grinded – repolished; #6: ARD – sintered – veneered; #7: control: analogous to #3 but without thermal cycling (TC) and mechanical loading (ML). FPDs were adhesively bonded to polymethylmethacrylate abutment teeth. TCML (TC: 6000 × 5°/55°; ML: 1.2 × 10⁶ × 50 N, 1.6 Hz) was conducted in a chewing simulator with steatite spheres as antagonists. Failures were monitored and fracture resistance was determined after ageing. Data were analysed statistically with Mann–Whitney U-test (Kolmogorov–Smirnov-test; α = 0.05). FPDs were subjected to scanning electron microscopy for fractographic failure analysis.ResultsNone of the FPDs failed during TCML, but showed wear at contact points. Median fracture force ranged between 1173.5 N (#4) and 1316.0 N (#3) without significant (p = 0.910) differences between the groups or in comparison to the control (p > 0.462).ConclusionsZirconia restorations showed high resistance to failures and fracture under different surface treatment variations. Full-contour polished or glazed zirconia FPDs might be an alternative to common veneered restorations.     

Effect of honey in preventing gingivitis and dental caries in patients undergoing orthodontic treatment

ObjectivesThis study was conducted to investigate the following: (1) the effects of chewing honey on plaque formation in orthodontic patients, (2) the effect of chewing honey on dental plaque bacterial counts, (3) determine if honey possesses antibacterial effects on bacteria recovered from plaques.MethodsFemale orthodontic patients (n = 20, 12–18 years of age) participated in this randomized controlled study. The effects of honey were compared to treatment with either 10% sucrose or 10% sorbitol that served as positive and negative controls, respectively. The pH of plaque was measured using a digital pH meter prior to baseline and at 2, 5, 10, 20, and 30 min after chewing honey or rinsing with control solutions and the numbers of Streptococcus mutans, Lactobacilli, and Prophymonas gingivalis in respective plaques were determined. The antibacterial activity of honey was tested against commonly used antibiotics using the disk diffusion method.ResultsSignificant differences in pH were observed in the honey and sucrose groups compared to the pH observed in the sorbitol group (p ? 0.001). The maximum pH drop occurred at 5 min in both the honey and sucrose groups; however the pH in the honey group rapidly recovered 10–20 min after exposure and did not drop below the critical decalcification pH of 5.5. On the other hand, the pH following sucrose exposure fell <5.5 and was associated with a 30 min recovery time. The pH observed for the sorbitol group did not change over time. Bacterial counts were significantly reduced in the honey group compared to the other treatment groups (p ? 0.001) and honey significantly inhibited the growth of all studied strains compared to inhibition observed with antibiotics (p ? 0.001).ConclusionsHoney can be used as an alternative to traditional remedies for the prevention of dental caries and gingivitis following orthodontic treatment.     

Osseointegration of fiber-reinforced composite implants: Histological and ultrastructural observations

ObjectivesThe aim of this study was to evaluate the bone tissue response to fiber-reinforced composite (FRC) in comparison with titanium (Ti) implants after 12 weeks of implantation in cancellous bone using histomorphometric and ultrastructural analysis.Materials and methodsThirty grit-blasted cylindrical FRC implants with BisGMA–TEGDMA polymer matrix were fabricated and divided into three groups: (1) 60 s light-cured FRC (FRC-L group), (2) 24 h polymerized FRC (FRC group), and (3) bioactive glass FRC (FRC–BAG group). Titanium implants were used as a control group. The surface analyses were performed with scanning electron microscopy and 3D SEM. The bone–implant contact (BIC) and bone area (BA) were determined using histomorphometry and SEM. Transmission electron microscopy (TEM) was performed on Focused Ion Beam prepared samples of the intact bone–implant interface.ResultsThe FRC, FRC–BAG and Ti implants were integrated into host bone. In contrast, FRC-L implants had a consistent fibrous capsule around the circumference of the entire implant separating the implant from direct bone contact. The highest values of BIC were obtained with FRC–BAG (58 ± 11%) and Ti implants (54 ± 13%), followed by FRC implants (48 ± 10%), but no significant differences in BIC or BA were observed (p = 0.07, p = 0.06, respectively). TEM images showed a direct contact between nanocrystalline hydroxyapatite of bone and both FRC and FRC–BAG surfaces.ConclusionFiber-reinforced composite implants are capable of establishing a close bone contact comparable with the osseointegration of titanium implants having similar surface roughness.     

Bone regeneration with BMP-2 and hydroxyapatite in critical-size calvarial defects in rats

BackgroundThis study describes the effect of bone formation by BMP-2 (bone morphogenetic protein-2), a bone formation inducer, with or without hydroxyapatite (HAP) application to critical-size defects in rat calvarial bone.Material and methodsTwenty male Wistar rats were divided into four groups of 5 animals each: control, HAP, BMP, and mixed BMP/HAP. A Critical-size defect of 4 mm was made using a trephine in the calvarial bone and, after that, BMP and/or HAP was applied to the defect according to the grouping. Defects were evaluated radiographically and histologically using ImageJ color analyzer software at 4 weeks postoperatively.ResultsThe histological data were more precise than the radiologic data due to the white color of the porous-type HAP material. The highest radiopacity was noted in the mixed BMP/HAP group (162.07 ± 9.06), followed by the HAP group (133.15 ± 21.8), then the BMP group (100.79 ± 8.27), and, lastly, the control group (54.45 ± 8.39). After subtracting the white background and using ImageJ for histological analysis, the highest rate of osteochondrogenesis was in the mixed BMP/HAP group (85.29% ± 8.21), and then the BMP group (77.34% ± 7.39), followed by the HAP group (59.82% ± 11.23), and, lastly, the control group (40.27% ± 7.44). Differences in the values between groups were then analyzed using confidence intervals (CI) of 95 and 99%.ConclusionWithin 4 weeks, the mixed BMP/HAP group showed the highest level of bone induction, especially compared to the BMP group, but this was non-significant; even with a 95% CI, the result was negative. This reveals that BMP alone can be applied, with a final result the same as that seen in the mixed BMP/HAP group. BMP and HAP, both being osteoinducting agents, even though they differ from a material classification point of view, have a positive effect on osteogenesis.     

Initial bone regeneration around fenestrated implants in Beagle dogs using basic fibroblast growth factor–gelatin hydrogel complex with varying biodegradation rates

PurposeThe purpose of this study was to compare the effectiveness of fast and slow biodegradation of basic fibroblast growth factor (bFGF)–gelatin hydrogel complex on bone regeneration around fenestrated implants as a new augmentation drug delivery system.MethodsNine titanium implants (3.3 mm diameter and 10 mm length) were placed into the edentulous areas of the mandibles of three adult beagle dogs with four screws exposed at the upper buccal side. The effectiveness of bFGF–gelatin hydrogel complexes of varying degradation types used to cover implant screws without membrane were compared with 1 μg and 10 μg bFGF–98 wt% gelatin as the fast degradation type and 10 μg bFGF–95 wt% gelatin as the slow degradation type. After 4 weeks, bone regeneration around the screws was evaluated histologically and histomorphometrically.ResultsWith use of 10 μg bFGF, regenerated bone around exposed screws was clearly seen in both the fast and slow degradation type groups. In contrast, little bone formation was seen in the fast degradation-type group with 1 μg bFGF. Height of regenerated bone for the slow degradation-type complex group was significantly greater than for the fast degradation-type group with 1 μg bFGF (P < 0.05).ConclusionThese results suggest that use of slow degradation-type bFGF–gelatin hydrogel complex may accelerate bone regeneration around fenestrated implants at an early stage of bone regeneration.     

The effect of rheumatoid arthritis and functional loading on the structure of the mandibular condyle in a transgenic mouse model: An FTIR study

ObjectiveThe objective of the present study was to investigate the effect of rheumatoid arthritis and functional loading through diet modification on the biochemical properties of the mandibular condyle in a transgenic mouse model and compare with healthy littermates.DesignTwenty three, 4-week old hybrid male mice were used. Eleven were of transgenic line hTNF 197 (Tg 197 – with rheumatoid arthritis – RA) and 12 healthy littermates, both from mixed background CBAxC57BL/6. Four groups of mice were formed. Group 1 [n = 5, RA-hard] included transgenic mice and received ordinary (hard) diet; group 2 [n = 6, RA-soft] included transgenic line and received soft diet; group 3 [n = 6, control-hard] were healthy littermates receiving ordinary (hard) diet and group 4 [n = 6, control-soft] were healthy littermates with soft diet. Experimental period was 28 days. Following sacrifice, the mandibular condyles were subjected to micro-attenuated reflection Fourier transform infrared spectroscopy (micro-ATR FTIR) to reveal collagen/proteoglycan conformation of the condylar cartilage, while resin-embedded and metallographically polished specimens were evaluated through reflection FTIR microscopy to identify mineralization status of the corresponding condylar bone.ResultsThe multivariable analysis revealed significantly lower a-helix to amide I percentage area ratio for the transgenic animals after adjusting for diet (β = −4.29, 95% CIs: −8.52, −0.06; p = 0.04). Mineral phase indices did not differ significantly between RA and control groups regardless the type of diet.ConclusionsInternal derangement of the anatomical structure with denaturation in the collagen structural components of the mandibular condyles of the RA animals was found, while no association with functional loading through diet modification was recorded.     

BMP2 and BMP4 variations and risk of non-syndromic cleft lip and palate

ObjectiveNon-syndromic cleft lip with or without cleft palate (CL/P) is one of the most common congenital anomalies and arises from the interaction of environmental and genetic factors. The objective of this study was to investigate the association between the BMP2 (bone morphogenetic protein 2) and BMP4 (bone morphogenetic protein 4) polymorphisms with non-syndromic CL/P to clarify the potential role of these genes in the etiology of CL/P in Iranian population.DesignThe allelic and genotypic frequencies of BMP2 rs235768 A > T and BMP4 rs17563 T > C polymorphisms were determined in 107 unrelated Iranian subjects with non-syndromic CL/P and 186 control subjects using PCR and RFLP methods, and the results were compared with healthy controls. A p-value of <0.05 was considered statistically significant.ResultsThe BMP2 rs235768 AT genotype was significantly higher (P = 0.009, OR = 3, 95% CI = 1.3–7.0) in the CL/P (59.8%) than the control group (33.3%). Similarly, the BMP4 rs17563 TC genotype were significantly higher (P = 0.008, OR = 3.7, 95% CI = 1.4–9.9) in the CL/P (70.0%) than the control group (44.6%).ConclusionThe BMP2 rs235768 A > T and BMP4 rs17563 T > C polymorphisms could be considered as the risk factor for non-syndromic CL/P in Iranian population.     

Adiponectin prevents orthodontic tooth movement in rats

ObjectiveThe objective was to examine the effects of repetitive local administration of adiponectin on experimental tooth movement in rats.Materials and methodsThe maxillary right first molar of male Wistar rats (n = 24) was moved mesially for 14 days, with local adiponectin injections (0.2 or 2 μg) every third day. Micro-computed tomography was performed at days 0, 6 and 14 and molar movement, bone density and bone volume fraction were calculated from the scans. Changes in extracellular matrix collagen and cell numbers in the periodontal ligament were analyzed histologically, and levels of circulating cytokines were measured by Luminex and ELISA.ResultsAdiponectin injections induced a reduction in tooth movement after 12 and 14 days compared to controls. No tooth movement was observed between days 3 and 14 in the group receiving the highest dosage (2 μg) of adiponectin. Differences in bone density and bone volume fractions between treatment and control groups were not identified. Relative size and morphology of collagen fibrils, and cell number in the periodontal region after adiponectin injections were unchanged compared to controls. Levels of circulating adiponectin or other selected factors in plasma were not influenced by the adiponectin injections.ConclusionsSubmucosal injections of adiponectin prevented experimental tooth movement in rats. The effect was dosage-dependent and local. Adiponectin injections caused no detectable changes in bone density, periodontal cell number or collagen content.