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To use resonance frequency analysis to evaluate tapered implants placed at maxillary posterior sites after lateral sinus augmentation.Patients who had missing teeth in the maxillary posterior area and required lateral sinus augmentation before implant placement were enrolled in this study. After a 6-month healing period, a tapered implant (Osstem TSIV) was placed. Implant success rate, survival rate, and marginal bone loss of the implants were measured. For resonance frequency analysis, implant stability quotient (ISQ) values were measured at each visit during a 1.5-year follow-up period.Twenty-four patients completed the study procedure. The residual bone height was 2.57 ± 1.10 mm (mean ± SD). Healing of the grafted area was uneventful in all cases, and 55 tapered implants were installed. The implant success rate was 95.56%, and the survival rate was 100% throughout the observation period. The marginal bone loss was limited to 0.22 ± 0.44 mm. ISQ increased gradually from 68.40 ± 11.14 to 82.24 ± 4.75 during the 1.5-year follow-up period.The tapered implants showed good initial and final stability after placement in the soft bone of the maxillary posterior area after lateral sinus augmentation.

This study aimed to quantify the healing following vertical augmentation of allogenic bone blocks with/without recombinant human bone morphogenetic protein-2 (rhBMP-2) on rabbit calvaria. Experiments were performed using allogenic bone blocks which were grafted bilaterally with or without rhBMP-2 on 20 rabbit calvaria, and these animals were divided to four groups according to the use of rhBMP-2 and healing periods (2 and 8 weeks; n = 10 in each group). Onlay-type bone blocks (8 mm in diameter and 5 mm high) were fixed with a self-tapping screw after removing the cortex in the control group, and the same protocol was applied with the addition of soaking the bone blocks with rhBMP-2 for 15 min in the test group. Radiographic and histologic analyses were performed after 2 or 8 weeks to evaluate the volumetric stability and bone regeneration within the grafted area. The radiographic analysis revealed that the height of the allogenic bone block decreased but its volume was maintained from 2 to 8 weeks in both the control and test groups. The histologic results demonstrated a statistically significant increase in new bone area in the test group, especially in the lower region adjacent to the preexisting calvarial floor. The amount of newly formed bone in all regions of the augmented bone blocks in both the control and test groups was greater at 8 weeks than at 2 weeks. In conclusion, the vertically grafted allogenic bone block maintained its volume with new bone formation, and this was accelerated by the addition of rhBMP-2. These findings indicate that allogenic bone block soaked with rhBMP-2 could be a useful candidate biomaterial for vertical augmentation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2700-2707, 2018.

Background Various biomaterials have been introduced for vertical ridge augmentation to replace autogenous block bone grafting. Purpose This retrospective study radiographically evaluated dimensional alterations of the vertically augmented alveolar ridge using collagen membrane and 3 types of materials: autogenous bone block, allogenous bone block, and particulated bone substitute. Materials and methods The electronic medical records of 32 patients who received vertical ridge augmentation using 3 types of materials were searched: 9 for autogenous bone block, 12 for allogenous bone block, and 11 for particulated bone substitutes. The vertical bone gain, progression of bone resorption, and peri-implant marginal bone loss after prosthetic loading were measured on follow-up radiographs. Results The alveolar ridge was vertically augmented by 5.13 ± 1.61, 4.54 ± 2.48, and 3.90 ± 0.85 mm (mean ± standard deviation) after grafting with autogenous bone block, allogenous bone block, and particulated bone substitute, respectively. The radiographic vertical height of the augmented ridge that received autogenous bone block reduced continuously during the first year but was stable thereafter. Sites that received allogenous bone block or particulated bone substitute exhibited dimensional shrinkage for up to 1.5 years postsurgery. However, the peri-implant marginal bone loss did not exceed 1 mm throughout the observational periods in all groups. Conclusions The clinical findings of this study suggest that the alveolar ridge can be vertically augmented using either allogenous bone block or particulated bone substitute. However, they require a longer healing period to ensure dimensional stability compared to the autogenous bone block.

Background Deproteinized porcine bone mineral (DPBM) was recently developed and commercially available in maxillary sinus grafting, in which demineralized bovine bone mineral (DBBM) was widely used. Objectives The present randomized controlled clinical trial aimed to compare histological bone quality and radiographic volume stability in maxillary sinuses grafted with DPBM and DBBM. Materials and Methods Twenty sinuses in 16 participants were enrolled and randomly allocated to control and test groups using sequentially numbered, sealed envelopes; laterally approached sinus grafting with DBBM and DPBM, respectively. All participants were blinded to the group assignment during the entire experiment. After standardized osteotomy at the lateral wall of the maxillary sinus, the sinus membrane was elevated, and the control or test biomaterial was grafted. Computed tomography (CT) images were taken immediately after surgery, and another CT and trephine biopsy was taken for radiographic and histological analyses after 6 months. The histological bone quality was measured as a primary outcome, and changes in the height and volume of the graft were evaluated in the reconstructed CT images as secondary outcomes. Results Fifteen sites (7 and 8 sites for control and test group) in 11 participants were finally included in the per protocol (PP) analysis, and 16 sites (7 and 9 sites, respectively) in 12 participants were included in the intention-to-treat (ITT) analysis; there were four drop-outs and one minor protocol violation. In both statistical analyses, the test groups showed comparable new bone formation and residual biomaterials in histology, and both groups exhibited minimal volume/height changes in radiographies. However, smaller sizes of residual biomaterials were observed in the histological samples from the test compared to control sites, despite the use of the same sizes of both biomaterials. Conclusions The results suggested that DPBM might produce comparable bone formation and volumetric stability with DBBM in maxillary sinus grafting, however, further clinical study with longer-term periods and larger sample sizes should be needed for confirming this suggestion.

OBJECTIVES The aim of this retrospective study was to determine the clinical and the radiographic outcomes of dental implants placed in elderly people older than 65 years. MATERIALS AND METHODS In total, 902 implants in 346 patients (age: 65-89 years) were followed up for 2-17 years following the implant surgery. The survival rate of these implants was recorded and analyzed. Changes in marginal bone levels were also analyzed in serial radiographs, and Cox regression analysis for implant loss was performed. RESULTS The survival rates were 95.39% and 99.98% in the implant- and patient-based analyses, respectively (involving a total of 29 implant failures), and the marginal bone loss at the implants was 0.17 ± 0.71 mm (mean ± SD). The number of failures was greatest in patients aged 65-69 years. The Cox regression with shared frailty analysis showed that implant loss was significantly greater in those aged 65-69 years than in those aged 70-74 years (P

Background Despite accumulating evidence for the longitudinal stability of the marginal bone level around an implant, there is limited evidence of predisposing risk factors for marginal bone loss based on some implants in a relatively large patient population. Purpose The aim of this study was to retrospectively determine the marginal bone loss around Straumann tissue-level dental implants during follow-up periods among which the maximum lasts up to 10 years, as well as the predisposing risk factors for peri-implant marginal bone loss. Materials and methods This study analyzed 1692 Straumann tissue-level dental implants in 881 patients, and relevant data were collected. The peri-implant marginal bone level was measured on periodic radiographs, and the changes in bone level were analyzed cumulatively from surgery until up to 10 years later. The log-rank test was used to select candidate critical risk factors for marginal bone loss, and multivariate analysis using Cox regression with the shared frailty model was performed. Results The overall peri-implant bone loss was 0.07 ± 0.21mm, 0.09 ± 0.26mm, 0.14 ± 0.41mm, and 0.17 ± 0.45mm at 3, 5, 7, and 9 years, respectively. Only 14 implants showed pathologic marginal bone loss exceeding 2mm during the follow-up period. While 2 implants were removed with continuous progressive marginal bone loss, 5 of the 14 implants showed early bone loss exceeding 1mm within the first year but then subsequently tended to show a stable marginal bone level. In the other seven implants, bone loss started after the first year and progressed continuously. Multivariate analysis revealed that diameter of the implant affected the peri-implant marginal bone loss. Conclusions Straumann tissue-level dental implants showed only slight peri-implant marginal bone loss, with a very low incidence of pathologic marginal bone loss exceeding 2mm.

Background Implant dentistry is progressing based on extensive scientific results including preclinical and clinical studies. Researchers and clinicians have focused on implant designs and surface characteristics, which has resulted in various features being developed and introduced for enhancing osseointegration and reducing complications. Purpose The purpose of this study was to determine the cumulative survival rates of Straumann tissue-level dental implants over a 10-year period and identify the patterns of implant loss at a single research institution. Materials and methods In total, 1692 implants were installed in 881 patients who visited the Department of Periodontology, Dental Hospital, Yonsei University, Seoul from January 2003 to December 2009. Cases in which the implant was completely removed were defined as implant failures. Electronic or paper charts and radiographs were used to determine whether the implants failed. The survival rate of implants was analyzed using lifetime tables and Kaplan-Meier survival estimates. Log-rank test and Cox regression with shared frailty were used to analyze the risk factors and the types of implant failure. Results The 10-year cumulative survival rates were 98.23% and 95.70% at the implant and patient levels, respectively. Before installing a prosthesis (defined as the early stage), 13 implants in 10 patients were removed, while eight implants in seven patients were removed after completing a prosthesis (defined as the late stage). The cumulative survival rate was related to the implant diameter, length, site, and insertion torque. Most implant failures within 1 year were attributable to osseointegration failure. There were several cases of failure in the late stage without apparent marginal bone loss. Conclusion Straumann tissue-level dental implants showed low failure rates and can be considered a useful long-term treatment option. The length, placement site, and insertion torque might affect implant survival.

Human bone marrow mesenchymal stem cells (hBMSCs) were isolated from bone marrow of the vertebral body. The hBMSCs were cultured under either hypoxic (1% O2) or normoxic (21% O2; control) conditions and the characteristics as mesenchymal stem cells were compared. Results revealed that hypoxia reduced proliferative potential and colony-forming efficiency of hBMSCs, and significantly enhanced osteogenic and chondrogenic differentiation. The hBMSCs enhanced the regenerative potential of bone in vivo. In vitro synthesis of soluble and insoluble collagen was significantly increased in the hypoxic condition. In vivo collagen tissue regeneration was also enhanced under the hypoxic condition, with concomitant increased expressions of various subtypes of collagen and lysyl-oxidase family mRNA. MicroRNA assays revealed that miR-155-5p, which negatively regulates HIF-1α, was significantly highly expressed. These observations demonstrate that hBMSCs obtained from human vertebrae exhibit altered characteristics under hypoxic conditions, and each factor contributing to hBMSC-mediated tissue healing should be evaluated with the goal of allowing their clinical application.

Objectives The aim of this study was to evaluate HA coated with different ratios of TCP as a carrier for hABMSCs obtained during implant osteotomy in comparison to slowly-resorbing biomaterial, Bio-Oss, as a negative control, using in vitro and in vivo experiments. Materials and Methods Human ABMSCs (hABMSCs) harvested during implant osteotomy were transplanted using HA/TCP or Bio-Oss as carriers in a murine ectopic transplantation model (n = 12). Pore size and cell affinity were evaluated in vitro. The area of newly formed bone was analyzed histometrically, the number of osteocytes was counted, and immunohistochemical staining was conducted against several markers of osteogenesis, including alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), and osteopontin (OPN). Osteoclast formation was evaluated by tartrate-resistant acid phosphatase staining. Results The carrier materials had comparable pore sizes. The cell affinity assay resulted in a high proportion of cell adhesion (>90%) in all experimental groups. Substantial new bone and osteocyte formation was observed on both HA/TCP carriers, whereas it was minimal with Bio-Oss. Positive immunostaining for ALP, RUNX-2, OCN, and OPN was observed with HA/TCP, but only limited expression of osteogenic markers with Bio-Oss. Conversely, there was a minimal osteoclast presence with Bio-Oss, but a significant presence of osteoclasts with both HA/TCP carriers. Conclusions Both types of scaffolds, BCP and Bio-Oss, showed high stem cell-carrying potential, but the in vivo healing patterns of their complexes with hABMSC could be affected by the microenvironment on the surfaces of the scaffolds. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2015.

The effects of fibroblast growth factor-2 (FGF-2) on collagen tissue regeneration by human bone marrow stem cells (hBMSCs) were investigated. hBMSCs were isolated from human vertebral body bone marrow during vertebral surgery and a population of hBMSCs with the characteristics of mesenchymal stem cells was observed. The FGF-2 treatment (5 ng/mL) affected on the colony-forming efficiency, proliferation, and in vitro differentiation of hBMSCs. Insoluble/soluble collagen and hydroxyproline synthesis was significantly enhanced in hBMSCs expanded with FGF-2 and the treatment of FGF-2 caused a reduction in the mRNA expression of collagen type I, but an increase of collagen types II and III along with lysyl oxidase family genes. Collagen formation was also examined using an in vivo assay model by transplanting hBMSCs into immunocompromised mice (n=4) and the histologic and immunohistochemical results revealed that significantly more collagen with a well-organized structure was formed by FGF-2-treated hBMSCs at 8 weeks posttransplantation (P

This study aimed to quantify the healing following vertical augmentation of allogenic bone blocks with/without recombinant human bone morphogenetic protein-2 (rhBMP-2) on rabbit calvaria. Experiments were performed using allogenic bone blocks which were grafted bilaterally with or without rhBMP-2 on 20 rabbit calvaria, and these animals were divided to four groups according to the use of rhBMP-2 and healing periods (2 and 8 weeks; n = 10 in each group). Onlay-type bone blocks (8 mm in diameter and 5 mm high) were fixed with a self-tapping screw after removing the cortex in the control group, and the same protocol was applied with the addition of soaking the bone blocks with rhBMP-2 for 15 min in the test group. Radiographic and histologic analyses were performed after 2 or 8 weeks to evaluate the volumetric stability and bone regeneration within the grafted area. The radiographic analysis revealed that the height of the allogenic bone block decreased but its volume was maintained from 2 to 8 weeks in both the control and test groups. The histologic results demonstrated a statistically significant increase in new bone area in the test group, especially in the lower region adjacent to the preexisting calvarial floor. The amount of newly formed bone in all regions of the augmented bone blocks in both the control and test groups was greater at 8 weeks than at 2 weeks. In conclusion, the vertically grafted allogenic bone block maintained its volume with new bone formation, and this was accelerated by the addition of rhBMP-2. These findings indicate that allogenic bone block soaked with rhBMP-2 could be a useful candidate biomaterial for vertical augmentation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2700-2707, 2018.

Purpose The aim of the present randomized controlled trial was to determine the safety and efficacy of injectable demineralized bone matrix (DBM) gel combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) on alveolar ridge preservation after tooth extraction. Materials and Methods A total of 69 patients were randomly assigned to either a test group (n = 35) or a control group (n = 34). In the test group, DBM, together with rhBMP-2 (0.05 mg/mL; rhBMP-2/DBM) was transplanted into the extraction sockets. The control group received DBM alone. The safety of rhBMP-2/DBM was evaluated by oral examination, serum chemistry, and hematologic examination. The radiographic changes in alveolar bone height and width were measured using computed tomography scans performed immediately after transplant and again 3 months thereafter. Results Healing was uneventful in all subjects, with no anticipated adverse events and no clinically significant changes in the serum chemistry and hematologic findings. No meaningful immune response was found among the study groups. No significant difference was found in the radiographic changes of alveolar bone height and width ( P > .05). Conclusions This new injectable biomaterial can be used easily and safely in clinical applications.

There is growing concern about unwanted effects associated with the clinical use of recombinant human bone morphogenetic protein-2 (rhBMP-2) at high concentrations, including cyst-like bone formation and excessive fatty marrow formation. We, therefore, evaluated the induction of mineralized/adipose tissue formation and the bone-healing pattern associated with the controlled release of E. coli-derived rhBMP-2 (ErhBMP-2) by a heparin-conjugated fibrin (HCF) system using ectopic and orthotopic in vivo models, respectively. In the ectopic transplantation model, mineralized tissue formed at the most superficial layer of the transplanted area and on the surfaces of grafted materials, and most of the interstitial space within the transplanted area was filled with excessive adipose tissue specifically at sites that received ErhBMP-2. However, sites that received ErhBMP-2 and HCF showed significantly increased mineralized tissue formation and decreased adipose tissue formation compared to the normal fibrin system with ErhBMP-2. In the orthotopic (calvarial defect) model, controlled release of ErhBMP-2 induced by HCF significantly reduced adipose tissue formation within the defect area compared to the clinically approved absorbable collagen sponge. From these results, it can be concluded that the use of a HCF system loaded with ErhBMP-2 may reduce adipose tissue formation and enhance mineralized tissue formation. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 545–554, 2015.

Purpose Various surgical techniques target achieving adequate keratinized tissue around dental implants; however, these techniques are usually performed before implant placement or upon the exposure of submerged implants. The aim of this case report is to describe a simultaneous placement of an interpositional free gingival graft (iFGG) with that of nonsubmerged implants in a patient lacking keratinized tissue and to assess the long-term outcome of this grafted gingiva. Methods A wedge-shaped free gingnival graft (FGG), including an epithelium-connective tissue (E-C) portion and a connective-tissue-only (CT) portion, was harvested from the palate. The CT portion was inserted under the buccal flap, and the E-C portion was secured tightly around the implants and to the lingual flap. Results At the 8-year follow-up, the gingival graft remained firmly attached and was well maintained, with no conspicuous shrinkage or reported discomfort during oral hygiene procedures. The use of an iFGG at a nonsubmerged implant placement minimizes the required number of surgical steps and patient discomfort while providing adequate buccal keratinized tissue. Conclusions Therefore, the technique could be considered an alternative method in increasing the keratinized tissue for cases that have a minimal amount of keratinized tissue. Graphical Abstract

Block-type biphasic calcium phosphate (BCP) carriers are more effective at delivering recombinant human bone morphogenetic protein-2 (rhBMP-2) in various clinical situations than are particle-type carriers, due to their potential for highly successful three-dimensional bone regeneration. The aim of this study was to confirm the bone-regenerative capabilities of three-dimensional BCP blocks with a low hydroxyapatite/β-tricalcium phosphate ratio (20/80) combined with collagen (10% wt) as an rhBMP-2 delivery system in a craniofacial vertical bone augmentation model. BCP blocks and BCP-collagen blocks (with average macropore sizes of 296 and 390 μm, respectively) with or without rhBMP-2 were fixed with osteosynthesis screws to the calvarial surface of rabbits. After 8 weeks, histologic and histomorphometric analyses were performed to evaluate the resulting new bone area, augmented area, bone density, and degree of integration. The area of new bone was significantly greater in specimens containing rhBMP-2 than in the control group (p < 0.05). Moreover, the area fractions of newly formed bone within the augmented area and a degree of integration between the regenerative bone and the calvarium were both significantly greater in the BCP-collagen/rhBMP-2 group than in the BCP/rhBMP-2 group (p < 0.05), whereas the two carrier systems exhibited similar rhBMP-2 release profiles, with sustained and linear release. The BCP and BCP/rhBMP-2 blocks exhibited excellent structural integrity, with large fragments of residual ceramic. In conclusion, the BCP-collagen composite block exhibited enhanced osteoinductive potential and could be a good candidate as a carrier of rhBMP-2 due to its characteristics of favorable volumetric stability, ease of handling, and excellent remodeling properties.

Purpose Mandibular tori are located above the mylohyoid ridge. These features are relatively common in most populations, but their prevalence varies with race. The purpose of this study was to identify the prevalence and features of mandibular tori in a Korean population using computed tomographic (CT) images. Materials and Methods The CT images of 726 patients were analyzed in this study. The presence or absence of a mandibular torus was identified on the CT images of each patient's quadrants and categorized according to gender and age. The most common location was also evaluated according to the corresponding teeth. The cortical thickness of each torus was evaluated using a 3-dimensional reconstruction software program. Results The prevalence of mandibular tori in the entire sample was 24.1%, and most cases were bilateral. Mandibular tori were observed more frequently in young adults, with the prevalence being inversely related to age. The most frequently observed location was the premolar region, and the mean cortical thickness was 6.9 mm. Conclusions Mandibular tori seem to change with aging. The prevalence in the present sample composed of Koreans was 24.1%, and younger patients may be more likely to develop a mandibular torus. Most tori presented a medium size bilaterally in the lingual aspect of the premolar region, and all were composed of cortical bone.

Park J-C, Kim J.C, Kim B-K, Cho K-S, Im G-I, Kim B-S, Kim C-S. Dose- and time-dependent effects of recombinant human bone morphogenetic protein-2 on the osteogenic and adipogenic potentials of alveolar bone-derived stromal cells. J Periodont Res 2012; 47: 645–654. © 2012 John Wiley & Sons A/S Background and Objective: Recombinant human bone morphogenetic protein-2 (rhBMP-2) is a well-known growth factor that can induce robust bone formation, and recent studies have shown that rhBMP-2-induced osteogenesis is closely related to adipogenesis. The aim of the present study was to determine the dose- and time-dependent effects of rhBMP-2 on the osteogenic and adipogenic differentiation of human alveolar bone-derived stromal cells (hABCs) in vivo and in vitro. Material and Methods: hABCs were isolated and cultured, and then transplanted using a carrier treated either with or without rhBMP-2 (100 μg/mL) into an ectopic subcutaneous mouse model. Comprehensive histologic and histometric analyses were performed after an 8-wk healing period. To further understand the dose-dependent (0, 10, 50, 200, 500 and 1000 ng/mL) and time-dependent (0, 3, 5, 7 and 14 d) effects of rhBMP-2 on osteogenic and adipogenic differentiation, in vitro osteogenic and adipogenic differentiation of hABCs were evaluated, and the expression of related mRNAs, including those for alkaline phosphatase, osteocalcin, bone sialoprotein, peroxisome-proliferator-activated receptor gamma-2 and lipoprotein lipase, were assessed using quantitative RT-PCR. Results: rhBMP-2 significantly promoted the osteogenic and adipogenic differentiation of hABCs in vivo, and gradually increased both the osteogenic and adipogenic potential in a dose- and time-dependent manner with minimal deviation in vitro. The expression of osteogenesis- and adipogenesis-associated mRNAs were concomitantly up-regulated by rhBMP-2. Conclusion: The findings of the present study showed that rhBMP-2 significantly enhanced the adipogenic as well as the osteogenic potential of hABCs in dose- and time-dependent manner. The control of adipogenic differentiation of hABCs should be considered when regenerating the alveolar bone using rhBMP-2.

Objectives Implant osteotomy yields a substantial amount of bone in the form of bone chips entrapped within drill flutes, and can provide a promising cell source for tissue engineering. The aims of this study were to isolate human alveolar bone-derived stromal cells (hABCs) obtained during implant osteotomy, and to evaluate osteogenic differentiation capacity of hABCs. Material and methods Bone chips were obtained by minimally irrigated implant drilling technique from 10 human donors. Isolated cells were studied with respect to their colony-forming efficiency, surface marker expression by immunofluorescence staining, fluorescence-activated cell sorting analysis and self-renewal potency. To verify the differentiation activity, in vitro osteogenic and adipogenic gene expressions were evaluated by reverse transcription-polymerase chain reaction, and in vitro formation of mineralized nodule and adipocytes was also evaluated. In vivo bone-forming activity was assessed by ectopic transplantation in immunocompromised mice (n = 5). Results Human alveolar bone-derived stromal cells population with characteristics of mesenchymal stem cells was present in the isolated cells. Upon hABC transplantation, significant ectopic bone formation was induced with the characteristics of fully matured bone tissue. Conclusion The data support the feasibility of using hABCs as a source of stem cells for dentoalveolar bone tissue reconstruction. The cell source has an advantage that the hABCs can be easily acquired during implant surgery.

Kim Y-T, Park J-C, Choi S-H, Cho K-S, Im G-I, Kim B-S, Kim C-S. The dynamic healing profile of human periodontal ligament stem cells: histological and immunohistochemical analysis using an ectopic transplantation model. J Periodont Res 2012; 47: 514–524. © 2012 John Wiley & Sons A/S Background and Objective: Human periodontal ligament stem cells (hPDLSCs) have been reported to play the pivotal role in periodontal regeneration. However, the dynamic cellular healing process initiated by hPDLSCs still remains to be elucidated. In the present study, the sequence of regeneration by hPDLSCs was assessed using histological and immunohistochemical observation in an ectopic transplantation model, which is a well-standardized assessment tool that excludes the innate healing factors from the animals. Material and Methods: Human periodontal ligament stem cells that were isolated and characterized from teeth (n = 12) extracted for the purpose of orthodontic treatment were transplanted with carriers into ectopic subcutaneous pouches in immunocompromised mice (n = 20). Animals were killed after several different healing periods: 3 d (n = 4), 1 (n = 4), 2 (n = 4), 4 (n = 4) and 8 wk (n = 4). Histological analysis for regenerated tissues formed by hPDLSCs was conducted using hematoxylin and eosin, Masson’s trichrome and picrosirius red staining. In addition, immunohistochemical staining was performed to observe the sequential expression of osteogenic/cementogenic and periodontal ligament tissue-specific markers associated with periodontal regeneration. Results: The whole healing process by transplanted hPDLSCs could be broadly divided into four distinctive phases. In the first phase, proliferated hPDLSCs migrated evenly all over the carrier, and collagenous tissues appeared in the form of amorphous collagen matrices. In the second phase, collagen fibers were well arranged among the carriers, and cementoid-like tissues were observed. In the third phase, the formation of mature collagen fibers, resembling Sharpey’s fibers, was associated with active mineralization of cementum-like tissues, and in the fourth phase, the maturation of cementum-like tissues was observed on carrier surfaces. Various osteogenic/cementogenic markers related to the regeneration processes were expressed in a well-orchestrated time order. Interestingly, well-organized cementum-like and periodontal ligament fiber-like tissues and cells with early and late osteogenic/cementogenic markers were frequently observed in the secluded area of carrier surfaces. We termed this area the cell-rich zone. Conclusion: The results from this study clearly demonstrated the sequential histological changes during periodontal tissue regeneration by hPDLSCs. Understanding of this process would potentially enable us to develop better cell-based treatment techniques.

Oral Diseases (2012) 18, 537–547 Objective: The human periodontal ligament stem cells (hPDLSCs) and human alveolar bone–derived stromal cells (hABCs) seem to be closely involved in the maintenance of alveolar bone in an anatomically indirect manner; however, there is little study on this matter. Therefore, the effect of hPDLSCs on the osteoclastogenic, osteogenic, and adipogenic differentiation of hABCs was evaluated, focusing on the humoral factors released by hPDLSCs. Materials and methods: Human periodontal ligament stem cells and hABCs were isolated and characterized. hPDLSCs were indirectly cocultured to observe the in vitro effect of humoral factors released from hPDLSCs on the osteoclastogenic, osteogenic, and adipogenic differentiation of hABCs. Human gingival fibroblasts (hGFs) were utilized as positive control. Results: Isolated cells demonstrated the presence of stem cells within. Indirect coculture of hPDLSCs greatly inhibited osteoclastogenesis by hABCs. Osteogenesis/adipogenesis of hABCs was also inhibited by indirect coculture with hPDLSC. The magnitude of regulatory effect from hPDLSCs was significantly greater than that of hGFs. Conclusions: Humoral factors released from hPDLSCs seemed to modulate the differentiation of hABCs, and the osteoclastogenic, osteogenic, and adipogenic differentiation of hABCs was all inhibited, suggesting the potential role of hPDLSCs in the maintenance of the alveolar bone.

Purpose: There has been no attempt to establish an objective implant surgical evaluation protocol to assess residents’ surgical competence and improve their surgical outcomes. The present study presents a newly developed assessment and rating system and simulation model that can assist the teaching staffs to evaluate the surgical events and surgical skills of residents objectively. Methods: Articles published in peer-reviewed English journals were selected using several scientific databases and subsequently reviewed regarding surgical competence and assessment tools. Particularly, medical journals reporting rating and evaluation protocols for various types of medical surgeries were thoroughly analyzed. Based on these studies, an implant surgical technique assessment and rating system (iSTAR) has been developed. Also, a specialized dental typodont was developed for the valid and reliable assessment of surgery. Results: The iSTAR consists of two parts including surgical information and task-specific checklists. Specialized simulation model was subsequently produced and can be used in combination with iSTAR. Conclusions: The assessment and rating system provided may serve as a reference guide for teaching staffs to evaluate the residents’ implant surgical techniques.

Bone morphogenetic proteins (BMP), in particular BMP-2, are the growth factors primarily responsible for osteoinduction. A knowledge of interactions between bone substitute materials and growth factor variants is crucial to designing bone substitutes with an ideal release profile. Here we compare glycosylated and non-glycosylated recombinant human bone morphogenetic protein-2 (rhBMP-2) either incorporated into a hydrolyzable polyethylene glycol (PEG) hydrogel developed as a slow release system or adsorbed to a deproteinized bovine bone matrix (DBBM), a clinically well-established bone substitute material. rhBMP-2 loaded materials were immersed in cell culture medium and rhBMP-2 concentration profiles in the supernatant were determined by an enzyme-linked immunosorbent assay. The corresponding biological activities were assessed in vitro by alkaline phosphatase activity assay. We show a strong affinity of rhBMP-2 for DBBM and reduced biological activity after its release from PEG hydrogels. Glycosylated rhBMP-2 was significantly less affected by the hydrogel and interacted significantly more strongly with DBBM than non-glycosylated rhBMP-2. We therefore question the combination of PEG hydrogels with DBBM as a rhBMP-2 delivery system over DBBM alone, since rhBMP-2 released from the hydrogel will be trapped by DBBM. Moreover, our results suggest that glycosylated rhBMP-2 is favorable in combination with PEG hydrogels, since its activity is better preserved, whereas in combination with DBBM non-glycosylated rhBMP-2 is favorable, benefiting from an initially higher concentration of free rhBMP-2.

Purpose The aim of this study was to elucidate the effect of a bovine hydroxyapatite/collagen (BHC) block in one-wall intrabony periodontal defects in dogs. Methods A one-wall intrabony periodontal defect (4 mm wide and 5 mm deep) was prepared bilaterally at the mesial side of the mandibular fourth premolar in five beagle dogs. After thorough root planing, block-type BHC (4×5×5 mm) was placed on one side. The contralateral defect area did not receive any material as a sham-surgery control. Histological analysis of the sites was performed after an 8-week healing period. Results Two of five samples in the experimental group healed well without dissipation of the graft materials, and histological analysis revealed excellent regeneration of the periodontal tissues. However, most of the grafted materials had been displaced in the other three samples, leaving only a small portion of the graft. The measured parameters exhibited large standard deviations, and the mean values did not differ significantly between the experimental and sham-surgery control sides. Conclusions The application of BHC alone-without a barrier membrane-to wide, one-wall intrabony periodontal defects yielded inconsistent results regarding both periodontal regeneration and substantivity of the graft materials. Thus, the use of a barrier membrane for noncontained-type defects is recommended to improve the stability of the grafted material, and to condense it.

Park J-C, So S-S, Jung I-H, Yun J-H, Choi S-H, Cho K-S, Kim C-S. Induction of bone formation by Escherichia coli-expressed recombinant human bone morphogenetic protein-2 using block-type macroporous biphasic calcium phosphate in orthotopic and ectopic rat models. J Periodont Res 2011; 46: 682–690. © 2011 John Wiley & Sons A/S Background and Objective: The potential of the Escherichia coli-expressed recombinant human bone morphogenetic protein-2 (ErhBMP-2) to support new bone formation/maturation using a block-type of macroporous biphasic calcium phosphate (bMBCP) carrier was evaluated in an orthotopic and ectopic rat model. Material and Methods: Critical-size (Φ 8 mm) calvarial defects and subcutaneous pockets in 32 Sprague–Dawley rats received implants of rhBMP-2 (2.5 μg) in a bMBCP carrier or bMBCP alone (control). Implant sites were evaluated using histological and histometric analysis following 2- and 8-wk healing intervals (eight animals/group/interval). Results: ErhBMP-2/bMBCP supported significantly greater bone formation at 2 and 8 wk (10.8% and 25.4%, respectively) than the control at 2 and 8 wk (5.3% and 14.0%, respectively) in calvarial defects (p

Park J-C, Kim J-M, Jung I-H, Kim JC, Choi S-H, Cho K-S, Kim C-S. Isolation and characterization of human periodontal ligament (PDL) stem cells (PDLSCs) from the inflamed PDL tissue: in vitro and in vivo evaluations. J Clin Periodontol 2011; 38: 721–731. doi: 10.1111/j.1600-051X.2011.01716.x. Abstract Objectives: Mesenchymal stem cells (MSC) could be isolated from healthy periodontal ligaments (PDL). The aims of this study were to isolate and characterize human PDL stem cells (hPDLSCs) from inflamed PDL tissue, and to evaluate their regenerative potential. Materials and Methods: Inflamed hPDLSCs (ihPDLSCs) were isolated from the inflamed PDL tissue obtained from intra-bony defects during flap surgery, and characterized by immunohistochemical staining, colony-forming unit assay, fluorescence-activated cell sorting, and mRNA expression in comparison with healthy hPDLSCs obtained from extracted teeth for orthodontic purpose. The proliferative potential and migratory potential was evaluated, and compared with healthy hPDLSCs. Regenerative potential was assessed by an in vivo ectopic transplantation model. Results: ihPDLSCs were successfully isolated and characterized as MSCs. Both ihPDLSCs and hPDLSCs were successfully differentiated under osteogenic/cementogenic and adipogenic microenvironment. The proliferative potential did not differ between healthy hPDLSCs and ihPDLSCs, while the migratory capacity was significantly increased in ihPDLSCs (p

Objectives The aim of this study was to evaluate the osteogenic and space-providing effect of rhBMP-2 produced in an Escherichia coli expression system (ErhBMP-2) with absorbable collagen block (ACB) as the carrier system in the rat calvarial defect model. Study design Eight-millimeter diameter calvarial defects were created in 60 male Sprague-Dawley rats. The animals were divided into 6 groups containing 10 animals each that received sham-surgery control (no material applied), ACB control (ACB alone), or ErhBMP-2/ACB (ACB loaded with 0.025 mg/mL ErhBMP-2). Histological and histometric analysis was performed after 2- and 8-week healing intervals. Results On histological observation, the level of bone formation in the defects was generally higher at 8 weeks than at 2 weeks. Surgical implantation of ErhBMP-2/ACB resulted in the enhanced bone regeneration compared with controls. Moreover, the collagen remnants of ACB appeared to be completely resorbed in ACB control and ErhBMP-2/ACB group after 8 weeks. Histometic analysis revealed that the ErhBMP-2/ACB group had a significantly greater augmented area, new bone area, and new bone ratio after 2 and 8 weeks than both control groups. However, there was no difference between 2 and 8 weeks in the ErhBMP-2/ACB group in all aspects of the augmented area, new bone area, and new bone ratio ( P Conclusion ErhBMP-2 loaded on ACB can induce favorable bone formation in the rat calvarial defect model, but this collagen carrier in block type did not fulfill the space-maintaining expectations for bone formation by ErhBMP-2.

Oral Diseases (2010) 17, 26–32 Objective: This study aimed to evaluate the validity of a surgically created interproximal periodontal defect in dogs. Materials and methods: Surgery was performed in the interproximal area between the maxillary second and third premolars in two beagle dogs. Following an incision and reflection of the gingival flap, a 3-mm wide and 5-mm high defect was prepared surgically at the interproximal area. A thorough root planing was performed and the flap was coronally positioned and sutured. The contra-lateral area was served as the control with no surgical intervention. After 8 weeks of healing, the animals were killed and the defect was analysed histometrically and radiographically. Results: The interproximal periodontal defect resembled a naturally occurring defect and mimicked a clinical situation. After healing, the defect showed limited bone (0.89 ± 0.02 mm) and cementum regeneration (1.50 ± 0.48 mm). Conclusions: Within the limitations of this pilot study, the interproximal periodontal defect showed limited bone and cementum regeneration. Thus, it can be considered as a standardized, reproducible defect model for testing new biomaterials.

Purpose: There is no consensus regarding the relationship between the width of keratinized mucosa and the health of periimplant tissues, but clinicians prefer to provide enough keratinized mucosa around dental implants for long-term implant maintenance. An apically positioned flap during second stage implant surgery is the chosen method of widening the keratinized zone in simple procedures. However, the routine suture techniques used with this method tend to apply tension over the provisional abutments and decrease pre-existing keratinized mucosa. To overcome this shortcoming, a pre-fabricated implant-retained stent was designed to apply vertical pressure on the labial flap and stabilize it in a bucco-apical direction to create a wide keratinized mucous zone. Methods: During second stage implant surgery, an apically displaced, partial thickness flap with a lingualized incision was retracted. A pre-fabricated stent was clipped over the abutments after connecting to the provisional abutment. Vertical pressure was applied to displace the labial flap. No suture was required and the stent was removed after 10 days. Results: A clinically relevant amount of keratinized mucosa was achieved around the dental implants. Buccally displaced keratinized mucosa was firmly attached to the underlying periosteum. A slight shrinkage of the keratinized zone was noted after the healing period in one patient, but no discomfort during oral hygiene was reported. Clinically healthy gingiva with enough keratinized mucosa was achieved in both patients. Conclusions: The proposed technique is a simple and time-effective technique for preserving and providing keratinized tissue around dental implants

The present study evaluated the effect of anodized surface implants and combined surface implants with calcium phosphate (CaP) coating on gap defect healing in dogs. The anodized + coated group showed higher bone-to-implant contact (BIC) and bone density (BD) than controls (anodized only) or the sandblasted/acid-etched (SLA) + anodized + coated group. The SLA + anodized + coated group showed lower BIC than the anodized + coated group and lower BD than anodized + coated or control groups. Anodized + coated implants showed good osseointegration in surgically created gap healing, but SLA + anodized + coated implants appeared less predictable in achieving osseointegration because of the offset effects of their advantages.

Recombinant human growth/differentiation factor-5 (rhGDF-5) is being evaluated as a candidate therapy in support of periodontal regeneration. The objective of this study was to evaluate cementum and alveolar bone formation, and aberrant healing events following surgical implantation of rhGDF-5 in an absorbable collagen sponge (ACS) carrier using an established periodontal defect model.Bilateral 4 x 5 mm (width x depth), one-wall, critical-size, intrabony periodontal defects were surgically created at the mandibular second and fourth pre-molar teeth in 15 Beagle dogs. Five animals received 1 microg/defect and five animals 20 microg/defect rhGDF-5 in unilateral defect sites. Contralateral sites received treatments reported elsewhere. Five animals received rhGDF-5/ACS with 0 (buffer control) and 100 microg/defect rhGDF-5 in contralateral defect sites. The animals were euthanized at 8 weeks post-surgery for histologic and histometric evaluation.Surgical implantation of rhGDF-5 stimulated significant periodontal regeneration. Cementum formation was significantly enhanced in sites implanted with rhGDF-5 (1 and 100 microg) compared with control (p0.05). Similarly, bone formation height was significantly greater in sites receiving rhGDF-5 (1 and 100 microg) compared with control (p0.05). There were no significant or remarkable differences in bone and cementum formation within the selected dose interval (1, 20 and 100 microg rhGDF-5). None of the control or the rhGDF-5 sites exhibited root resorption, ankylosis, or other aberrant tissue reactions.Surgical implantation of rhGDF-5/ACS may be used safely to support periodontal wound healing/regeneration in intrabony periodontal defects without complications.

The purpose of this study was to evaluate the effect of post heat treatment on HA coated implants in healing of circumferential bone defects and to compare the results with those of bone graft treatment. The results presented higher bone to implant contact in both post heat treatment group and bone graft group than no heat treatment group and the controls, with no significance between the groups. The post heat treated HA coated implants seemed to show better osseointegration with bone and produce beneficial effect in resolving the defects as comparable with bone graft treatment.

Objectives: This study evaluated both the clinical and histological aspects of bone formation in maxillary sinus augmentation using MBCP as the bone-grafting material. Material and methods: MBCP was used as a primary bone substitute for maxillary sinus augmentation. Fifty-two patients were selected after a medical and dental examination, and were divided into the following three groups: those augmented with MBCP only; MBCP combined with irradiated cancellous bone; and MBCP combined with intraoral autogenous bone. After a healing period (average 6.78 months after surgery), bone cores were harvested for a histological evaluation and the implant fixtures were installed. These bone cores were evaluated via light microscope and implants were followed up for at least six months after loading. Results: Four to ten months after surgery, new vital bone surrounding the MBCP particles was observed in 18 bone biopsies. Two out of the 130 implants installed were explanted due to a failure of osseointegration before the prosthetic procedure. All the remaining implants were functioning for 6 to 27 months (average 12.96 months). The cumulative survival rate of the implants was 98.46%. Conclusion: These results show that MBCP can be used as a grafting material for sinus floor augmentation, whether combined with other bone graft materials or not, and lead to a predictable prognosis for dental implants in the posterior maxillary area where there is insufficient vertical height for fixture installation.

Calcium sulfate used in this study is newly fabricated to overcome its disadvantages; brittleness, difficult handling properties, rapid resorption and reaction heating. Especially when calcium sulfate is used in the graft procedure, it is difficult to control the setting time, solubility into blood and body fluid. To improve the handling properties, Calcium sulfate is mixed with Carboxylmethylcellulose (CMS). The putty type calcium sulfate used in the present study presented lower values in cementum formation and new bone formation than the control group. This may be due to use of poor osteoinductive calcium sulfate alone in the defect, limiting new bone formation. In addition, complete resorption of calcium sulfate after 8 weeks presents that addition of polymer failed to delay the resorption rate of calcium sulfate. Rather, it seemed to disturb the natural healing of the defect when compared with the findings in the control group. But the favorable result of calcium sulfate is shown when used with alloplast. The material used in the present study was developed for easier formability and manipulation avoiding scattering or dissolving of the material with blood or body fluid. However, the results reveal that further studies on effect of calcium sulfate with various other bone graft materials and ideal ratios in mixing the two should be determined in the future.However, the results reveal that further studies on effect of calcium sulfate with various other bone graft materials and ideal ratios in mixing the two should be determined in the future.

Periodontal plastic surgery is used to fulfill the esthetic and functional demands of patients. The palatal masticatory mucosa is the main donor site for connective tissue, and the thickness of the graft tissue obtained is an important factor for the success of this technique. The aim of this study was to measure the thickness of masticatory mucosa in the posterior palatal area using computerized tomography (CT).The thickness measurements were performed on the images of 100 adult subjects who underwent CT on the maxilla for implant surgery. Twenty-four standard measurement points were defined in the hard palate according to the gingival margin and the middle palatal suture. The radiographic measurements were used after calibration. The data were analyzed to determine the differences in the mucosal thickness according to gender, age, tooth position, and depth of the palatal vault.The overall mean thickness of the palatal masticatory mucosa was 3.83 +/- 0.58 mm (range: 2.29 to 6.25 mm). Females had significantly thinner mean masticatory mucosa (3.66 +/- 0.52 mm) than males (3.95 +/- 0.60 mm) (P0.0001). The thickness of the palatal masticatory mucosa increased with age. The mean thickness according to tooth site was 3.46 mm (maxillary canine), 3.66 mm (first premolar), 3.81 mm (second premolar), 3.13 mm (first molar), 3.31 mm (the base of the interproximal papilla of the first and second molars), and 3.39 mm (second molar). There was an overall increase in the thickness of the palatal masticatory mucosa as the distance from the gingival margin to the middle palatine suture increased, with the exception of the Ca-d (a point at 12 mm from the gingival margin of the canine) region. There was no significant difference in the thickness of the palatal masticatory mucosa between the groups with high or low palatal vaults.The palatal masticatory mucosa thickness increased from the canine to premolar region but decreased at the first molar region and increased again in the second molar region, with the thinnest area at the first molar region and the thickest at the second premolar region. The canine to premolar region seems to be the most appropriate donor site that contains a uniformly thick mucosa. CT can be considered an alternative method for the measurement of palatal soft tissue thickness.

This study investigated the bone growth pattern in surgically created coronal defects with various depths around implants in dogs.Four mongrel dogs were used. All mandibular premolars were extracted under general anesthesia and left to heal for 2 months. After ostectomy, bony defects were prepared in test sites, using a stepped drill with a diameter of 6.3 mm and two depths: 2.5 mm (test sites 1 [T1]) and 5.0 mm (test sites 2 [T2]). In the control sites, the implants were placed after ostectomy without any coronal defects. T1, T2, and control sites were prepared in the right and left sides of the mandible. Six implants, 3.3 mm in diameter and 10 mm in length, were placed in each dog; the implants were submerged completely. Two dogs were sacrificed 8 weeks after surgery, and the other two dogs were sacrificed 12 weeks after surgery. The stability of all implants was measured with a resonance frequency analyzer after placement and after sacrifice. All sites were block-dissected for ground sectioning and histologic examination.After 12 weeks of healing, only T2 were not filled fully with bone. At week 8, the mean bone-to-implant contact (BIC) was 47.7% for control, 43.6% for T1, and 22.2% for T2. At week 12, the control BIC was 56.7% and the 2.5-mm defect had a greater BIC (58.8%). However, in the 5-mm defect, the BIC was 35.1%. At insertion, stability was reduced at sites with a greater defect depth. Similar stability was noted in all specimens after 8 and 12 weeks of healing.Bone healing between an implant and marginal bone was compromised at sites with a deeper defect when the width of the bone defect was 1.5 mm.

The development and expanded use of dental implants has been remarkably increased. The implant stability and osseointegration are important factors in the success of treatment. Commercially pure titanium and its alloys are the most commonly used materials for endosseous dental implants. Hydroxyapatite (HA), one of the calcium phosphate, has been studied for biomedical applications since it has similarity in chemistry with inorganic components of human bone and its biocompatibility. But the use of HA coated dental implants remain still controversial, because some reports suggest that the HA coating may separate from the substructure, undergo dissolution in tissue fluids, and contribute to rapid breakdown around the implants. HA coating can be applied on metal implants by numerous methods. Of several coating methods, ion beam-assisted deposition (IBAD) has shown to promise. This study was to compare the healing response of various nano-coating surface implants (by IBAD) placed in surgically created circumferential gaps 12 weeks postplacement. In six mongrel dogs, all mandible premolars and the first molar were extracted and after an 8-week healing period, four kinds of implants were placed. 2 mm circumferential coronal defects around the implants were performed surgically with a customized step drill. Groups were divided according to the fixture surfaces. The dogs were sacrificed following a 12 week healing period. Specimens were analyzed histologically and histomorphometrically. During the healing period, healing was uneventful and implants were well maintained. The bone-to-implant contact (BIC) for experimental groups —calcium phosphate coating implants with or without heat treatment- were much higher than the control group. The defect fill were also higher in the experimental groups. In the control group, the coronal part showed minimal bone fill and osseointegration. Within the scope of this study, calcium phosphate coating using IBAD may improve the bone response and the calcium phosphate coating may therefore be suitable in implant design with complex surface geometries. Copyright © 2008 John Wiley & Sons, Ltd.

The purpose of this study was to evaluate the effects of a chitosan membrane coated with polylactic and polyglycolic acid (PLGA) on bone regeneration in a rat calvarial defect. Surgical implantation of chitosan membranes resulted in enhanced local bone formation at both 2 and 8 weeks. In conclusion, the chitosan membrane coated with PLGA had a significant potential to induce bone formation in the rat calvarial defect model. Within the selected PLGA dose range and observation intervals, there appeared to be no meaningful differences in bone formation.

The purpose of this study was to evaluate the regenerative effects of a tetracycline blended polylactic and polyglycolic acid (TC-PLGA) and non-blended polylactic and polyglycolic acid (PLGA) barrier membrane on one-wall intrabony defects in beagle dogs. It can be concluded that when used for guided tissue regeneration TC-PLGA membranes show a beneficial effect on one-wall intrabony defects in beagle dogs.

The aim of this study was to investigate the application of electrochemical technology in relation to the electrolysis of water in specific fields, including mouthrinses for the control of periodontal disease and the cleaning of contact lenses. A platinum (Pt) coated titanium (Ti) electrode cell (Pt/Ti cell, Dolki Korea Ltd.) was used for electrochemical experiments. Using the disinfection or oxidation potential of the Pt/Ti cell, sterilized water was prepared for clinical testing. An experimental group used mouthrinses with the sterilized water, where as the control group did not. In the experimental group, the gingival and plaque indices, the probing pocket depth and clinical attachment level all showed statistically significant decreases, but in the control group showed significant increases (P

The effects of hydroxyapatite (HA)-coating onto collagen carriers for application of recombinant human bone morphogenetic protein 2 (rhBMP-2) on cell differentiation in vitro, and on in vivo healing patterns after sinus-augmentation and alveolar socket-grafting were evaluated. In vitro induction of osteogenic/adipogenic differentiation was compared between the culture media with rhBMP-2 solution and with the released rhBMP-2 from the control collagen and from the HA-coated collagen. Demineralized bovine bone and collagen/HA-coated collagen were grafted with/without rhBMP-2 in sinus-augmentation and tooth-extraction-socket models. Adipogenic induction by rhBMP-2 released from HA-coated collagen was significantly reduced compared to collagen. In the sinus-augmentation model, sites that received rhBMP-2 exhibited large amounts of vascular tissue formation at two weeks and increased adipose tissue formation at eight weeks, this could be significantly reduced by using HA-coated collagen as a carrier for rhBMP-2. In extraction-socket grafting, dimensional reduction of alveolar ridge was significantly decreased at sites received rhBMP-2 compared to control sites, but adipose tissue was increased within the regenerated socket area. In conclusion, HA-coated collagen carrier for Escherichia coli-derived rhBMP-2 (ErhBMP-2) may reduce in vitro induction of adipogenic differentiation and in vivo adipose bone marrow tissue formation in bone tissue engineering by ErhBMP-2.

OBJECTIVES This study evaluated the dimensional ridge alteration in a buccal-bone-deficient extraction socket, and ridge regeneration following socket grafting accompanied by recombinant human bone morphogenetic protein 2 (rhBMP-2) or a collagen membrane covering. MATERIAL AND METHODS In five beagle dogs, entire buccal bone of the extracted sockets of premolars was surgically removed and immediately grafted using one of the following graft protocols: (1) sham surgery without any grafting, and grafting with (2) deproteinized bovine bone mineral (DBBM), (3) DBBM/rhBMP-2 and (4) DBBM covered with a collagen membrane (DBBM/Membrane). Quantitative/qualitative analyses were performed radiographically/histologically after 8 weeks. RESULTS Buccal-deficient extraction sockets healed with significant reduction in buccolingual dimension along the entire length of the socket, but all grafting techniques reduced the dimensional changes compared to the non-grafted control sites. Histologically, sites received DBBM only exhibited minimal regeneration, whereas sites grafted with DBBM/rhBMP-2 or DBBM/Membrane exhibited greater new bone formation extending the entire augmented area. CONCLUSIONS Buccal-bone-deficiency may lead to significant volume reduction after tooth extraction along the entire length of the socket, and socket grafting accompanied by rhBMP-2 or covered with a membrane can be candidate therapies for preservation of the buccolingual dimension and successful ridge regeneration.

Beta tricalcium phosphate (beta-TCP) has been developed as one of the carriers of recombinant human bone morphogenetic protein (rhBMP). However, it is not known whether the particle size of beta-TCP is related to its resorption rate and the degree of bone formation. The purpose of this study was to evaluate the effect of using beta-TCP with different particle sizes on the ability of rhBMP-4 to enhance bone formation in the rat calvarial defect model.Calvarial, 8-mm-diameter, critical-size defects were created in 100 male Sprague-Dawley rats. Five groups of 20 animals each received either rhBMP-4 (2.5 microg) using beta-TCP with a particle size of 50 to 150 microm, rhBMP-4 (2.5 microg) using beta-TCP with a particle size of 150 to 500 microm, a beta-TCP control with a particle size of 50 to 150 microm, a beta-TCP control with a particle size of 150 to 500 microm, or a sham-surgery control, respectively, and were evaluated by measuring their histologic and histometric parameters following a 2- and 8-week healing interval.There were no significant differences in the defect closure, new bone area, or augmented area between either the two rhBMP-4/beta-TCP groups or between the two beta-TCP control groups at 2 and 8 weeks.rhBMP-4 combined with either small- or large-particle beta-TCP had a significant effect on the induction of bone formation compared to either a small- or large-particle beta-TCP control or a sham-surgery control. Within the parameters of this study, varying the particle size of beta-TCP did not seem to have a significant effect on bone formation.

Dehiscence bone defects, frequently observed on dental implants placed in periodontitis-affected alveolar bone or extraction sockets were treated with β-tricalcium phosphate (β –TCP) and chitosan membrane for guided bone regeneration, and the new bone formation on the treated sites were studied. Beagle dogs were used for the experiment. First to fourth mandibular premolars were extracted, and the post extraction alveolar bone surface was planed. After 8 weeks of healing, 3 by 4mm dehiscence defects were created using straight fissure burs. Total of 16 oxidized titanium surface implants were placed on the bone defects of the subjects, two on each side. Control sites were treated with implants only. Experimental Group 1 sites were treated with implants and chitosan membrane. Experimental Group 2 sites were treated with implants, β-TCP and chitosan membrane. Experimental Group 3 sites were treated with implants, β-TCP, autogenous bone and chitosan membrane. The animals were sacrificed 12 weeks after implant placement, and the specimens from the treated sites were histologically studied with following results. Limited amount of new bone formation was observed in control group with unexposed membrane. Slightly greater amount of bone formation was observed on sites treated with β-TCP+membrane or autogenous bone+ β-TCP+membrane compared to control group. Remnants of chitosan membrane and β-TCP encapsulated with connective tissue were observed during experimental periods. These results suggest that further studies are needed on membrane rigidity and infection control for space maintenance underneath the membrane and bone substitutes in the treatment of dehiscence defects.

This study evaluated periodontal repair and biomaterial reaction following implantation of a calcium carbonate and autogenous bone on the regeneration of 3-wall intrabony defects in the beagle dogs. The surgical control group received a flap operation only, while the experimental group I was treated with a calcium carbonate and the experimental group II was treated with autograft. The subjects were sacrificed 8 weeks after surgery and a comparative histometric analysis was done. No root resorption or ankylosis were observed in the experimental group. ANOVA post Hoc test showed that the experimental group II produced statistically significant higher gain in connective tissue adhesion, new cementum regeneration and new bone formation. These results suggest that the use of autograft in 3-wall intrabony defects has a significant effect on new cementum and new bone formation height.

Currently, more than 20 bone morphogenetic proteins (BMPs) have been identified, and many trials have been carried out using recombinant human BMPs (rhBMPs) for bone tissue engineering. However, comparative analyses on bone formative activities of rhBMP using a preclinical model have been limited. Therefore, the aim of this study was to evaluate and compare the osteogenic potential of rhBMP-2, -4, and -7 delivered with absorbable collagen sponge (ACS) upon early (2 weeks) and complete (8 weeks) wound healing phases in a critical sized rat calvarial defect model.Eight-millimeter critical sized calvarial defects were created in 30 male Sprague-Dawley rats. The animals were divided into three groups of 10 animals each. The defects were treated with 0.025 mg/ml rhBMP-2/ACS, rhBMP-4/ACS, or rhBMP-7/ACS. The rats were sacrificed at either 2 (five rats) or 8 (five rats) weeks after surgery, and the results were evaluated histologically, histomorphometrically, and immunohistometrically.The surgical implantation of rhBMP-2/ACS, rhBMP-4/ACS, or rhBMP-7/ACS resulted in enhanced local bone formation in the rat calvarial defect model at both 2 and 8 weeks. The amount of defect closure, new bone area, and bone density were similar in the three groups at each time point (P0.05). In terms of bone density and new bone area, there were statistically significant differences between results obtained at 2 weeks and those obtained at 8 weeks in all groups (P0.05). Two-way analysis of variance (ANOVA) revealed that there was no correlation between the time and conditions (P0.05), but time was found to have a strong influence on defect closure, new bone area, and bone density (P0.05). Irrespective of rhBMP type, positive immunoreactions of osteopontin (OPN) and osteocalcin (OCN) were evident at 2 and 8 weeks. Intense OPN and OCN staining was observed near the newly formed bone as well as in some cells within the new bone.Within the rhBMP types used, rhBMP concentration, and the observation interval, there appears to be no specific differences in bone regenerative potential. All rhBMPs used in this study may be considered effective factors for inducing bone formation.

Aim: Autogenous bone grafts and bone biomaterials are being used as part of protocols aiming at reconstruction of periodontal defects. There is a limited biologic information on the effect of such materials on periodontal healing, in particular aberrant healing events that may prevent their general use. The objective of this study was, using histological techniques, to evaluate periodontal healing with focus on root resorption and ankylosis following implantation of autogenous bone and a coral-derived biomaterial into intra-bony defects in dogs. Methods: One-wall intra-bony periodontal defects were surgically created at the distal aspect of the second and the mesial aspect of the fourth mandibular premolars in either right or left jaw quadrants in four Beagle dogs. Each animal received particulated autogenous bone and the resorbable calcium carbonate biomaterial into discrete one-wall intra-bony defects. The mucoperiosteal flaps were positioned and sutured to their pre-surgery position. The animals were euthanized 8 weeks post-surgery when block sections of the defect sites were collected and prepared for qualitative histological analysis. Results: There were no significant differences in periodontal healing between sites receiving autograft bone and the coral-derived biomaterial. A well-organized periodontal ligament bridging new bone and cementum regeneration was observed extending coronal to a notch prepared to delineate the apical extent of the defect. Osteoid and bone with enclosed osteocytes were formed onto the surface of both autograft and coral particles. Although small resorption pits were evident in most teeth, importantly none of the biomaterials provoked marked root resorption. Ankylosis was not observed. Conclusion: Particulated autogenous bone and the coral-derived biomaterial may be implanted into periodontal defects without significant healing aberrations such as root resorption and ankylosis. The histopathological evaluation suggests that the autogenous bone graft has a limited osteogenic potential as demonstrated in this study model.