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OBJECTIVE To test whether or not topically administered recombinant human epidermal growth factor (rhEGF) accelerates the early healing phase of oral soft tissue wounds. METHODS One day following the creation of palatal defects (n = 6/animal), 14 dogs were allocated to one of the following five groups: spontaneous healing (SH), vehicle ointment (V), vehicle ointment + rhEGF at concentrations of 1 μg/g (EGF1), 10 μg/g (EGF10) or 50 μg/g (EGF50). Topical administration of ointments was repeated twice per day until sacrifice at days 8 and 16. Wound area was clinically monitored. Keratinocytes proliferation (Ki67-immunolabelling), inflammatory response (IR) and areas of collagen (C) and granulation tissue (GT) were histologically measured. Kruskal-Wallis test with Dunnett correction was used for multiple group statistical comparisons. RESULTS Clinically, in comparison with SH, a significantly smaller wound area was observed in groups EGF1 and EGF10 at day 8 (p

Background This experimental study aims to evaluate the effect of hyaluronic acid on healing of infected extraction sockets compared with recombinant human bone morphogenetic protein-2 (rhBMP-2). Methods Both third and fourth mandibular premolars of six beagle dogs were hemisected, and the distal roots were extracted at baseline. Subsequently, combined endodontic-periodontic lesions were induced at the remaining mesial roots. After 4 months, the mesial roots on both sides of the mandible were removed. Four sockets per dog were randomly allocated to four groups: Group 1, Control; Group 2, only absorbable collagen sponge (ACS: carrier); Group 3, 1% hyaluronic acid (HA) gel + ACS; and Group 4, rhBMP-2 + ACS. After 3 months of healing, the dogs were euthanized for microcomputed tomography and histologic analysis. Results After the lesion induction period (4 months), communication between the periodontal lesion and endodontic periapical lesion was observed at all remaining mesial roots. Alveolar bone overgrowth was observed in groups 3 and 4, but bone volume density was not significantly different among all groups. At the crestal portion, mineralization, and osteocalcin expression were higher in groups 3 and 4 than in groups 1 and 2. Conclusion Treatment with HA can promote bone formation and improve the wound healing rate comparable to rhBMP-2 in infected extraction sockets.

There have been previous studies on the relationship between periodontitis and peri-implantitis, but limited information is available on how periodontitis affects osseointegration and wound healing of newly placed dental implants adjacent to natural teeth. The objective of the present experiment is to evaluate healing around dental implants adjacent to teeth with untreated experimental periodontitis.The study included six male beagle dogs. Scaling and plaque control procedures were performed on three dogs (control group). In the other three dogs (experimental group), retraction cords and ligature wires were placed subgingivally around all premolars and the first molars. Induced experimental periodontitis was confirmed after 3 months. Each control or experimental group was divided into two subgroups depending on the timing of implant placement (immediate/delayed). Twelve dental implants (two implants for each dog) were placed immediately, and the other 12 dental implants (two implants for each dog) were placed 2 months after extraction. The animals were sacrificed 2 months after implant placement. Histologic and histometric analyses were performed.Four implants (three from the immediate placement group and one from the delayed placement group) failed in the experimental group. There were significant differences in the percentage of bone-to-implant contact and marginal bone volume density between the control and experimental groups. Both parameters were significantly lower in the experimental group than in the control group (P0.05). There was a tendency toward more marginal bone loss in the experimental group than the control group.Immediate placement of implants is associated with a higher failure rate compared with delayed placement. Untreated experimental periodontitis was correlated with compromised osseointegration in the implants with delayed placement.

Background: Pain from local anesthetic injection makes patients anxious when visiting a dental clinic. This study aims to determine differences in pain according to types of local anesthetizing methods and to identify the possible contributing factors (e.g., dental anxiety, stress, and sex). Methods: Thirty-one patients who underwent open-flap debridement in maxillary premolar and molar areas during treatment for chronic periodontitis were evaluated for this study. A randomized, split-mouth, single-masked clinical trial was implemented. The dental anxiety scale (DAS) and perceived stress scale (PSS) were administered before surgery. Two lidocaine ampules for each patient were used for local infiltration anesthesia (supraperiosteal injection). Injection pain was measured immediately after local infiltration anesthesia using the visual analog pain scale (VAS) questionnaire. Results from the questionnaire were used to assess degree of pain patients feel when a conventional local anesthetic technique (CNV) is used compared with a computer-controlled anesthetic delivery system (CNR). Results: DAS and PSS did not correlate to injection pain. VAS scores were lower for CNR than for CNV regardless of the order in which anesthetic procedures were applied. VAS score did not differ significantly with sex. Pearson coefficient for correlation between VAS scores for the two procedures was 0.80, also indicating a strong correlation. Conclusion: Within the limitations of the present study, relief from injection pain is observed using CNR. J Periodontol 2016;87:783-789.

Peri-implantitis is a chronic inflammatory process with advanced bone loss and impaired healing potential. For peri-implantitis treatment, tissue engineering can be applied to enhance bone regeneration of peri-implant defects. This study aimed to evaluate ex vivo bone morphogenetic protein 2 (BMP2) gene delivery using canine periodontal ligament stem cells (PDLSCs) for regeneration of peri-implantitis defects. Canine PDLSCs were transduced with adenoviral vectors containing BMP2 (BMP2/PDLSCs). After peri-implantitis was induced by ligature placement in six beagle dogs, regenerative procedures were performed; hydroxyapatite (HA) particles and collagen gel with autologous canine PDLSCs (PDLSC group) or BMP2/PDLSCs (BMP/PDLSC group) or without cells (control group) were grafted into the defects and covered by an absorbable membrane. Three months later, the animals were sacrificed. In vitro, BMP2/PDLSCs showed similar levels of stem cell properties to PDLSCs, such as colony-forming efficiency and expression of MSC markers STRO-1 and CD 146. BMP2/PDLSCs produced BMP-2 until day 21 at a concentration of 4-8 ng/mL. In vivo, the BMP2/PDLSC group showed significantly more new bone formation and re-osseointegration in peri-implantitis defects compared to the other groups. In conclusion, ex vivo BMP2 gene delivery using PDLSCs enhanced new bone formation and re-osseointegration in peri-implantitis defects.

To the best of our knowledge, the influence of external versus internal implant-abutment connections on crestal bone remodeling has not been reported. The aim of the present study is to investigate the influence of the abutment connection on peri-implant crestal bone levels (CBLs) using radiographic recordings.Radiographic recordings from 40 single-tooth implants (20 external and 20 internal octagonal connections; one implant/patient) in 40 patients (15 males and 25 females; mean age: 54.3 years) were selected for analyses. The radiographic evaluation included the following: 1) linear bone change (LBC); 2) dimensional change (DC); and 3) angle between the implant and adjacent bone (AIB). Differences in LBC, DC, and AIB between implant placement and 1 year after loading for each system were evaluated using a paired t test. Comparison of LBC, DC, and AIB between systems at 1 year after loading was done using analysis of covariance. The significance level was set at P ≤0.05.Radiographic CBLs (LBCs) were reduced at 1 year after loading compared to those at implant placement to reach statistical significance for the external connection (P = 0.000) but not the internal connection (P = 0.939). CBL changes were significantly greater for the external compared to the internal connection (P = 0.000). Similarly, the DC for the external connection was significantly greater compared to that for the internal connection (P = 0.004).Within the limitations of this study, the implant-abutment connection technology appears to have a significant impact on peri-implant CBLs, with the external connection paralleled by a significant reduction of CBLs.

In previous studies, oligopeptide corresponding to the cell-binding domains of bone morphogenetic proteins that bind to bone morphogenetic protein receptor enhanced the bone regenerative capacity of bovine bone minerals (BBM). The aim of this study is to evaluate the ability of BBM coated with oligopeptide to promote periodontal regeneration in a 1-wall intrabony defect model in dogs.The second and third mandibular premolars and first molars of six adult beagles were extracted bilaterally, and the extraction sites were allowed to heal for 10 weeks. The 1-wall intrabony defects were prepared bilaterally on the mesial and distal side of the fourth mandibular premolars. Twenty-four intrabony defects were assigned to four treatment groups: 1) open flap debridement; 2) guided tissue regeneration (GTR); 3) GTR with a collagen membrane and BBM; and 4) GTR with a collagen membrane and BBM coated with the oligopeptide (Pep-BBM). The animals were sacrificed 10 weeks after surgery. For the histometric analysis, defect height, junctional epithelium migration, new cementum, new bone height, and new bone area were measured. New bone volume was measured using microcomputed tomography.Wound healing was generally uneventful. For junctional epithelium migration, the BBM and Pep-BBM groups exhibited mean (± SE) values of 0.53 ± 0.41 and 0.48 ± 0.30 mm, and for new cementum height, 1.71 ± 0.46 and 2.50 ± 2.00 mm, respectively. For junctional epithelium migration and cementum regeneration, there were no significant differences between the two groups. The mean (± SE) values of new bone height and new bone volume in the Pep-BBM group (3.88 ± 0.31 mm and 32.35% ± 9.60%) were significantly greater than the mean values for the BBM group (2.60 ± 0.41 mm and 20.56% ± 1.89%). For bone regeneration, the Pep-BBM group showed superior results compared to the BBM group with statistically significant differences.Through various parameters to evaluate periodontal regeneration, this oligopeptide coating influenced only the ability of BBM to promote bone regeneration in 1-wall intrabony defects in beagles. Junctional epithelium migration and cementum regeneration were not affected by this oligopeptide coating, and further investigations with special focus on regeneration of the periodontal ligament are necessary.

The aims of this study are to evaluate the efficacy of a eutectic mixture of local anesthetic (EMLA) cream on pain perception during scaling and to compare the intensities of pain provoked by hand and ultrasonic instruments.Forty subjects with chronic gingivitis or periodontitis were enrolled in the study. In this randomized, split-mouth, controlled, masked clinical trial, each of the four quadrants in subjects was randomly assigned to receive one of the following protocols: scaling with an ultrasonic scaler with or without the application of the EMLA cream or scaling with a Gracey curet with or without the application of the EMLA cream. Pain levels after each quadrant of scaling were assessed with a visual analog scale (VAS; 0 to 100 mm) and verbal rating score (VRS; 0 to 4). All subjects were recalled to detect any complications after 24 hours.The mean VAS and VRS when EMLA cream was applied (18.39 ± 14.47 mm and 0.95 ± 0.69) were significantly lower (P0.001 for VAS and VRS) compared to when EMLA cream was not used (26.54 ± 16.46 mm and 1.30 ± 0.75). The mean VAS and VRS of the ultrasonic scaler group (20.43 ± 16.40 mm and 0.98 ± 0.76) were significantly lower (P = 0.024 for VAS; P = 0.003 for VRS) than those of the Gracey curet group (24.50 ± 15.17 mm and 1.28 ± 0.69). None of the subjects showed adverse effects after EMLA-cream application.Although most patients experienced limited pain during scaling, a significant reduction of pain is achieved by using EMLA cream and ultrasonic scaler.

Our special thanks to Osstem Implant Inc., Busan, Korea for manufacturing the postulated drills that were used in this study.

To our knowledge, no study has evaluated the success or survival rate of single-tooth implants that replaced missing maxillary and mandibular second molars. The purpose of the present study was to evaluate the 1- to 5-year cumulative survival rate (CSR) for single-tooth implants placed in the second molar region and the effects of associated factors.Four hundred eighty-nine patients (298 males and 191 females; age range: 23 to 91 years; mean age: 47 years) who were admitted to the Seoul National University Dental Hospital between March 2003 and July 2008 and treated with single-tooth implants in the second molar region (227 maxillary implants and 294 mandibular implants; total: 521 implants) were included in the study. Thirty-two subjects received two implants each. A 1- to 5-year CSR was calculated using a life-table analysis. A comparison of CSRs between maxillary versus mandibular implants, one-stage versus two-stage implants, short (or=8.5 mm) versus long (10 mm) implants, and standard-diameter (or=4.0 mm) versus wide-diameter (or=5.0 mm) implants was performed using Wilcoxon (Gehan) statistics. P0.05 was considered significant.Fifteen of the 521 implants were lost between insertion and the follow-up examinations. The 1- to 5-year CSR was 95.1%. There were no statistically significant differences in CSRs between implants placed in maxillas and mandibles (96.3% versus 94.9%, respectively; P = 0.084), one- and two-stage implants (95.6% versus 94.7%, respectively; P = 0.267), short and long implants (100% versus 95.1%, respectively; P = 0.582), and standard- and wide-diameter implants (93.8% versus 96.8%, respectively; P = 0.065).Within the limitations of the study, the placement of single-tooth implants in the second molar region was an effective and reliable treatment modality. Also, associated factors such as implant diameter, length, and location (the maxilla versus the mandible) may not have an impact on the long-term success of implants.

Aim: The aim of the present study was to investigate bone regeneration following ex vivo bone morphogenetic protein-2 (BMP-2) gene delivery using human gingival fibroblasts (HGFs) in rat calvarial defects. Materials and Methods: An 8 mm craniotomy defect was created in Sprague― Dawley rats. The animals were divided into four groups: (1) non-grafted group, the defect was left empty; (2) collagen matrix group, the defect was filled with collagen matrix only; (3) HGF group, the defect was filled with non-transduced HGFs on collagen matrix; (4) BMP-2/HGF group, the defect was filled with BMP-2 gene-transduced HGFs on collagen matrix. Animals were sacrificed at 2 and 4 weeks after surgery, and micro-computed tomographic and histologic observations were performed. Results: The BMP-2/HGF group showed promoted osseous healing of calvarial defects, as compared with the other groups. At both 2 and 4 weeks, regenerated bone area was significantly greater in the BMP-2/HGF group than the other three groups. Quite a few number of transplanted HGFs were observed within the regenerated bone tissues. Conclusions: The results of this study suggest that ex vivo BMP-2 gene delivery induces prominent bone regeneration in vivo and HGFs may be useful as target cells for ex vivo gene therapy.

In a previous study, recombinant human transforming growth factor-beta1 (rhTGF-beta(1)) in a calcium carbonate carrier was implanted into critical-size, supraalveolar periodontal defects under conditions for guided tissue regeneration (GTR) to study whether rhTGF-beta(1) would enhance or accelerate periodontal regeneration. The results showed minimal benefits of rhTGF-beta(1), and a clear account for this could not be offered. One potential cause may be that the rhTGF-beta(1) formulation was biologically inactive. Several growth or differentiation factors have been suggested to accelerate degradation of biomaterials used as carriers. The objective of this study was to evaluate possible activity of rhTGF-beta(1) on biodegradation of the calcium carbonate carrier.rhTGF-beta(1) in a putty-formulated particulate calcium carbonate carrier was implanted into critical-size, supraalveolar periodontal defects under conditions for GTR in five beagle dogs. Contralateral defects received the calcium carbonate carrier combined with GTR without rhTGF-beta(1) (control). The animals were euthanized at week 4 post-surgery and block biopsies of the defect sites were collected for histologic and histometric analysis. Radiographs were obtained at defect creation and weeks 2 and 4 after defect creation.No statistically significant differences were observed in new bone formation (bone height and area) among the treatments. However, total residual carrier was significantly reduced in sites receiving rhTGF-beta(1) compared to control (P = 0.04). Similarly, carrier density was considerably reduced in sites receiving rhTGF-beta(1) compared to control; the difference was borderline statistically significant (P = 0.06).Within the limitations of the study, it may be concluded that rhTGF-beta(1) accelerates biodegradation of a particulate calcium carbonate biomaterial, indicating a biologic activity of the rhTGF-beta(1) formulation apparently not encompassing enhanced or accelerated periodontal regeneration.

Objectives: It has been suggested that degranulating platelet α-granules release growth factors having a potential to modulate bone formation. The objective of this study was to evaluate the osteoconductive potential of a platelet-rich plasma (PRP) preparation. Methods: Thirty adult male Sprague–Dawley rats were used. The PRP preparation was obtained from 10 ml of whole blood drawn from one age-matched donor rat. The preparation was processed by gradient density centrifugation and stored at −80°C until use. Using aseptic techniques, the PRP preparation soak loaded onto an absorbable collagen sponge (ACS) or ACS alone was surgically implanted into contralateral critical size 6-mm calvaria osteotomies in 18 animals. Twelve animals received ACS versus sham surgery in contralateral defects. Animals were sacrificed at 4 and 8 weeks when biopsies were collected for histologic and histometric analysis. Results: The animals were maintained without adverse events. Bone formation was highly variable in sites receiving PRP and control treatments. Defect bone fill at 4 weeks averaged (±SD) 28.8±27.4% (PRP/ACS) versus 39.1±24.4% (ACS; p=0.2626) and 62.0±20.0% (ACS) versus 71.6±32.2% (sham surgery; p=0.1088), and at 8 weeks 81.0±12.9% (PRP/ACS) versus 64.5±28.1% (ACS; p=0.2626) and 75.6±34.1% (ACS) versus 74.1±24.2% (sham surgery; p=0.7353). Remnants of the ACS biomaterial were observed at both 4 and 8 weeks in sites implanted with PRP/ACS or ACS. Conclusions: The results suggest that the PRP preparation has a limited potential to promote local bone formation.

Background: Traditionally, the mean value from histometric assessments of several histologic sections from the same site has been, used for the histometric characterization of the site. The histometric analysis of the supraalveolar periodontal defect model uses observations from buccal and lingual sites in three stepserial sections representing the central aspect of the mesial and distal root for the third and fourth mandibular premolar teeth. The objectives of this study were to investigate the correlations and determine whether there are significant differences in the measurement values of various histometric parameters between the three step-serial sections and to test the hypothesis that, in this animal model, histometric assessments using the most central section of each root are comparable to the mathematical means of corresponding assessments of the site using the three step-serial sections. Methods: Histometric analysis using light microscopy, an attached digital camera system, and a personal computer-based image analysis system including a custom program for the supraalveolar periodontal defect model was performed on histologic sections acquired from one jaw quadrant in each of 12 dogs. The animals had received a protocol including placement of a coral biomaterial and guided tissue regeneration (GTR) membranes, and were evaluated following a 4-week healing interval. Twelve parameters were assessed using three central step-serial sections from each root. For each parameter, pair-wise comparisons were performed using the mixed models analysis of variance, and the correlations between measurements were assessed by the intraclass correlation coefficient (ICC). Results: For all parameters assessed, there were no significant differences (P >0.05) among the three sections. The differences between the central section and the means of the three step-serial sections were small and not statistically significant. Parameters that could be assessed on most or all specimens, such as defect height and area, membrane height, and bone regeneration area, showed high correlations among the three step-serial sections (intraclass coefficient [ICC] ∼ 0.91 to 0.98), and between each of the three sections and the mean for the three step-serial sections (ICC ≥0.95). The most central section had somewhat higher correlation with the mean than measurements made on the lateral sections. Conclusion: The results suggest that using the supraalveolar periodontal defect model representative histometric data can be obtained evaluating only the central section from each defect site. J Periodontol 2004;75:1688-1693.

There is a limited understanding of the effect of bone biomaterials on the healing potential when used in conjunction with guided tissue regeneration (GTR). The objective of this study was to evaluate the effect of a space-providing coral-derived biomaterial on alveolar bone regeneration in conjunction with GTR.Bilateral, critical-size, 6-mm, supra-alveolar, periodontal defects were created in four young adult Beagle dogs. In a split-mouth design, the animals received an ePTFE device to provide for GTR in contralateral defect sites with or without the coral biomaterial. The animals were euthanized at 4 weeks post surgery. A histometric analysis assessed vertical regeneration of alveolar bone relative to space-provision by the ePTFE device. Because of the correlation of within-dog measurements, a mixed model ANOVA was used to analyze the data.There was significantly greater mean bone regeneration in sites receiving calcium carbonate coral implant GTR (cGTR) compared to GTR (p0.0001). Sites providing larger wound areas exhibited greater bone regeneration compared to sites exhibiting smaller wound areas (p0.0001). However, grouping the sites by wound area thresholds showed that bone regeneration was not significantly different in sites receiving cGTR compared to sites receiving GTR alone, irrespective of the size of the wound area (p0.5).Space-provision has a significant effect on bone regeneration following GTR. The coral biomaterial effectively enhances space-provision, and this appears to be the principal mechanism by which this biomaterial supports bone regeneration rather than postulated osteoconductive properties.

Background: Histometric assessments are routinely used to evaluate biologic events ascertained in histologic sections acquired from animal and human studies. The objective of this study was to evaluate the intra- and inter-examiner reproducibility of histometric assessments in the supraalveolar periodontal defect model. Methods: Histometric analysis using incandescent and polarized light microscopy, an attached digital camera system, and a PC-based image analysis system including a custom program for the supraalveolar periodontal defect model was performed on histologic sections acquired from one jaw quadrant in each of 12 dogs. The animals had received an experimental protocol including implantation of a coral biomaterial and guided tissue regeneration (GTR) barrier devices, and were evaluated following a 4-week healing interval. Histometric parameters were recorded and repeated within a 3-month interval by two examiners following brief training. Intra- and inter-examiner reproducibility was assessed using the intra-class correlation coefficient (ICC). Results: Most parameters showed high intra-examiner ICCs. Parameters including defect height, connective tissue repair, bone regeneration (height/area), formation of a junctional epithelium, positioning of the GTR device, ankylosis, root resorption, and defect area yielded an ICC0.9. The ICCs for bone density and biomaterial density were somewhat lower (0.8 and 0.7, respectively). The inter-examiner reproducibility was somewhat lower compared to the intra-examiner reproducibility. Nevertheless, the ICCs were generally high (ICC range: 0.6–0.9). Conclusions: Histometric evaluations in the supraalveolar periodontal defect model yield highly reproducible results, in particular when a single examiner performs the histometric measurements, even when the examiner was exposed to limited training.