Periodontal disease-associated compensatory expression of osteoprotegerin is lost in type 1 diabetes mellitus and correlates with alveolar bone destruction by regulating osteoclastogenesis

Alveolar bone resorption results from the inflammatory response to periodontal pathogens. Systemic diseases that affect the host response, such as type 1 diabetes mellitus (DM1), can potentiate the severity of periodontal disease (PD) and accelerate bone resorption. However, the biological mechanisms by which DM1 modulates PD are not fully understood. The aim of this study was to determine the influence of DM1 on alveolar bone resorption and to evaluate the role of receptor activator of nuclear factor-kappaB ligand (RANKL)/osteoprotegerin (OPG) in osteoclastogenesis in rats. PD was induced by means of ligature in nondiabetic and in streptozotocyn-induced DM1 rats. Morphological and morphometric analyses, stereology and osteoclast counting were performed. RANKL and OPG mRNA levels, protein content, and location were determined. PD caused alveolar bone resorption, increased the number of osteoclasts in the alveolar bone crest and also promoted changes in RANKL/OPG mRNA expression. DM1 alone showed alveolar bone destruction and an increased number of osteoclasts at the periapical and furcal regions. DM1 exacerbated these characteristics, with a greater impact on bone structure, resulting in a low OPG content and a higher RANKL/OPG ratio, which correlated with prominent osteoclastogenesis. This work demonstrates that the effects of PD and DM1 enhance bone destruction, confirms the importance of the RANKL signaling pathway in bone destruction in DM1 in animal models and suggests the existence of alternative mechanisms potentiating bone degradation in PD.

Correlation of Fractal Dimension with Histomorphometry in Maxillary SinuLifting Using Autogenous Bone Graft

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Reconstruction of the Alveolar Buccal Bone Plate in Compromised Fresh Socket after Immediate Implant Placement Followed by Immediate Provisionalization

ObjectiveThe aim of this clinical report was to reestablish the buccal bone wall after immediate implant placement. The socket defect was corrected with autogenous bone, and a connective tissue graft was removed from the maxillary tuberosity to increase the thickness, height, and width of the buccal bone and gingival tissue followed by immediate provisionalization of the crown during the same operation.Clinical ConsiderationsA 66-year-old patient presented with a hopeless maxillary left central incisor with loss of the buccal bone wall. Atraumatic, flapless extraction was performed, and an immediate implant was placed in the extraction socket followed by preparation of an immediate provisional restoration. Subsequently, immediate reconstruction of the buccal bone plate was performed, using the tuberosity as the donor site, to obtain block bone and connective tissue grafts, as well as particulate bone. Finally, immediate provisionalization of the crown followed by simple sutures was performed. Cone-beam computed tomography and periapical radiographs were taken before and after surgery. After 4 months, the final prosthetic crown was made. After a 2-year follow-up, a satisfactory aesthetic result was achieved with lower treatment time and morbidity.ConclusionThis case demonstrates the effective use of immediate reconstruction of the buccal bone wall for the treatment of a hopeless tooth in the maxillary aesthetic area. This procedure efficiently promoted harmonious gingival and bone architecture, recovered lost anatomical structures with sufficient width and thickness, and maintained the stability of the alveolar bone crest in a single procedure.Clinical SignificanceIf appropriate clinical conditions exist, immediate dentoalveolar restoration may be the most conservative means of reconstructing the buccal bone wall after immediate implant placement followed by immediate provisionalization with predictable healing and lower treatment time.

Diabetes reduces mesenchymal stem cells in fracture healing through a TNF alpha-mediated mechanism

Diabetes interferes with bone formation and impairs fracture healing, an important complication in humans and animal models. The aim of this study was to examine the impact of diabetes on mesenchymal stem cells (MSCs) during fracture repair.Fracture of the long bones was induced in a streptozotocin-induced type 1 diabetic mouse model with or without insulin or a specific TNF alpha inhibitor, pegsunercept. MSCs were detected with cluster designation-271 (also known as p75 neurotrophin receptor) or stem cell antigen-1 (Sca-1) antibodies in areas of new endochondral bone formation in the calluses. MSC apoptosis was measured by TUNEL assay and proliferation was measured by Ki67 antibody. In vitro apoptosis and proliferation were examined in C3H10T1/2 and human-bone-marrow-derived MSCs following transfection with FOXO1 small interfering (si)RNA.Diabetes significantly increased TNF alpha levels and reduced MSC numbers in new bone area. MSC numbers were restored to normal levels with insulin or pegsunercept treatment. Inhibition of TNF alpha significantly reduced MSC loss by increasing MSC proliferation and decreasing MSC apoptosis in diabetic animals, but had no effect on MSCs in normoglycaemic animals. In vitro experiments established that TNF alpha alone was sufficient to induce apoptosis and inhibit proliferation of MSCs. Furthermore, silencing forkhead box protein O1 (FOXO1) prevented TNF alpha-induced MSC apoptosis and reduced proliferation by regulating apoptotic and cell cycle genes.Diabetes-enhanced TNF alpha significantly reduced MSC numbers in new bone areas during fracture healing. Mechanistically, diabetes-enhanced TNF alpha reduced MSC proliferation and increased MSC apoptosis. Reducing the activity of TNF alpha in vivo may help to preserve endogenous MSCs and maximise regenerative potential in diabetic patients.

The effect of simvastatin treatment on bone repair of femoral fracture in animal model

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of bone marrow on osseointegration in long bones: an experimental study in sheep

Aim To evaluate the influence of yellow bone marrow on osseointegration of titanium oral implants using a long bone model.Material and methodsThe two tibiae of eight sheep were used as experimental sites. Two osteotomies for implant installation were prepared in each tibia. At the control sites, no further treatments were performed while, at the test sites, bone marrow was removed from the osteotomy site with a curette to an extent that exceeded the implant dimensions. As a result, the apical portion of the implants at the control sites was in contact with bone marrow while, at the test sites, it was in contact with the blood clot. After 2months, the same procedures were performed in the contralateral side. After another month, the animal was sacrificed. Ground sections were obtained for histological analysis.ResultsAfter 1month of healing, no differences between test and control sites were found in the apical extension of osseointegration and the percentage of new bone-to-implant contact. However, after 3months of healing, a higher percentage of new bone-to-implant contact was found at the test compared to the control sites in the marrow compartment. The apical extension of osseointegration, however, was similar to that found at the 1-month healing period both for test and control sites.ConclusionsOsseointegration appeared to be favored by the presence of a blood clot when compared to the presence of yellow fatty bone marrow. Moreover, the contact with cortical bone appeared to be a prerequisite for the osseointegration process in the long bone model.

Enamel matrix protein derivative plus synthetic bone substitute for the treatment of mandibular Class II furcation defects: a case series

Objectives: The aim of this study is to report on the treatment of mandibular Class II furcation defects with enamel matrix protein derivative (EMD) combined with a beta TCP/HA (beta-tricalcium phosphate/hydroxyapatite) alloplastic material. Method and Materials: Thirteen patients were selected. All patients were nonsmokers, systemically healthy, and diagnosed with chronic periodontitis; had not taken medications known to interfere with periodontal tissue health and healing; presented one Class II mandibular furcation defect with horizontal probing equal to or greater than 4 mm at buccal site. The clinical parameters evaluated were probing depth (PD), relative gingival margin position (RGMP), relative vertical clinical attachment level (RVCAL), and relative horizontal clinical attachment level (RHCAL). A paired Student t test was used to detect differences between the baseline and 6-month measurements, with the level of significance of .05. Results: After 6 months, the treatment produced a statistically significant reduction in PD and a significant gain in RVCAL and RHCAL, but no observable change in RGMP. RVCAL ranged from 13.77 (+/- 1.31) at baseline to 12.15 (+/- 1.29) after 6 months, with a mean change of -1.62 +/- 1.00 mm (P<.05). RHCAL ranged from 5.54 (+/- 0.75) to 2.92 (+/- 0.92), with a mean change of -2.62 +/- 0.63 mm (P<.05). After 6 months, 76.92% of the patients improved their diagnosis to Class I furcation defects while 23.08% remained as Class II. Conclusion: The present study has shown that positive clinical results may be expected from the combined treatment of Class II furcation defects with EMD and beta TCP/HA, especially considering the gain of horizontal attachment level. Despite this result, controlled clinical studies are needed to confirm our outcomes.

Morphological aspects of tympanic bulla after ventral osteotomy in cats

To evaluate tympanic bulla healing after experimental ventral osteotomy in cats. Twenty adult cats were submitted to unilateral ventral bulla osteotomy and divided into two groups: cats of A1 group (n=10) were euthanized at 8 weeks and cats of A2 group (n=10), at 16 weeks postoperative. Signs of Horner's syndrome or damage to the inner ear were not found. Open-mouth radiographs taken in the immediate postoperative showed interruption in the contour of the larger compartment of the operated bulla. The result of Mcnemar'test was significant in A2 group (*p=0.0156). Macroscopic exams revealed that the operated bullae were similar to the normal ones, with preservation of the tympanic cavity. Connective tissue at the osteotomy site of the larger compartment was significantly found in the operated bullae in both groups (McNemar test: A1 p=0.0020*; A2 p=0.0078*). Histomorphometric analyses showed that the connective tissue length at the osteotomy site was shorter in A2 group than in the A1 group (Mann-Whitney test: p=0.0021*). Experimental ventral osteotomy did not alter significantly the tympanic bulla conformation and complete regeneration of tympanic bulla frequently did not occur before 16 weeks of postoperative period.

Is bone transplantation the gold standard for repair of alveolar bone defects?

New strategies to fulfill craniofacial bone defects have gained attention in recent years due to the morbidity of autologous bone graft harvesting. We aimed to evaluate the in vivo efficacy of bone tissue engineering strategy using mesenchymal stem cells associated with two matrices (bovine bone mineral and α-tricalcium phosphate), compared to an autologous bone transfer. A total of 28 adult, male, non-immunosuppressed Wistar rats underwent a critical-sized osseous defect of 5 mm diameter in the alveolar region. Animals were divided into five groups. Group 1 (n = 7) defects were repaired with autogenous bone grafts; Group 2 (n = 5) defects were repaired with bovine bone mineral free of cells; Group 3 (n = 5) defects were repaired with bovine bone mineral loaded with mesenchymal stem cells; Group 4 (n = 5) defects were repaired with α-tricalcium phosphate free of cells; and Group 5 (n = 6) defects were repaired with α-tricalcium phosphate loaded with mesenchymal stem cells. Groups 2-5 were compared to Group 1, the reference group. Healing response was evaluated by histomorphometry and computerized tomography. Histomorphometrically, Group 1 showed 60.27% ± 16.13% of bone in the defect. Groups 2 and 3 showed 23.02% ± 8.6% (p = 0.01) and 38.35% ± 19.59% (p = 0.06) of bone in the defect, respectively. Groups 4 and 5 showed 51.48% ± 11.7% (p = 0.30) and 61.80% ± 2.14% (p = 0.88) of bone in the defect, respectively. Animals whose bone defects were repaired with α-tricalcium phosphate and mesenchymal stem cells presented the highest bone volume filling the defects; both were not statistically different from autogenous bone.

Use of autogenous bone and beta-tricalcium phosphate in maxillary sinus lifting: a prospective, randomized, volumetric computed tomography study

The correction of bone defects can be performed using autogenous or alloplastic materials, such as beta-tricalcium phosphate (β-TCP). This study compared the changes in bone volume (CBV) after maxillary sinus lifting using autogenous bone (n=12), autogenous bone associated with β-TCP 1:1 (ChronOS; DePuy Synthes, Paoli, CA, USA) (n=9), and β-TCP alone (n=11) as grafting material, by means of cone beam computed tomography (CBCT). CBV was evaluated by comparing CBCT scans obtained in the immediate postoperative period (5-7 days) and at 6 months postoperative in each group using OsiriX software (OsiriX Foundation, Geneva, Switzerland). The results showed an average resorption of 45.7±18.6% for the autogenous bone group, 43.8±18.4% for the autogenous bone+β-TCP group, and 38.3±16.6% for the β-TCP group. All bone substitute materials tested in this study presented satisfactory results for maxillary sinus lifting procedures regarding the maintenance of graft volume during the healing phase before the insertion of implants, as assessed by means of CBCT.

Influence of irrigation and intracanal dressing on the healing process of dogs' teeth with apical periodontitis.

Dogs' teeth with apical periodontitis were treated endodontically, Dakin's solution being used in an experimental group as the irrigation solution, and camphorated paramonochlorophenol as an intracanal dressing. For a second group of teeth, the irrigation solution used was physiologic saline, dry cotton only being placed into the pulpal chamber to take the place of an intracanal dressing. In a second visit, overinstrumentation and a new application of the same kind of dressing were performed, the root canals being then filled with gutta-percha cones and zinc oxide-eugenol cement. Other specimens were treated, in one visit, where physiologic saline or Dakin's solution were the irrigation solutions. The animals were sacrificed 6 months after the obturation of the root canals. Histologic exams showed better results for the experimental group where Dakin's solution and camphorated paramonochlorophenol had been used, with signs of repair characterized by newly formed cementum and bone tissue, as well as a marked reduction in size of the periapical lesions. No differences were seen in the results with physiologic saline or Dakin's solution as irrigants.

Influence of morphological variables in photoelastic models with implants submitted to axial loading

Purpose: This study used 12 photoelastics models with different height and thickness to evaluate if the axial loading of 100N on implants changes the morphology of the photoelastic reflection. Methods: For the photoelastic analysis, the models were placed in a reflection polariscope for observation of the isochromatic fringes patterns. The formation of these fringes resulted from an axial load of 100N applied to the midpoint of the healing abutment attached to the implant with 10.0mm x 3.75mm (Conexão, Sistemas de Próteses, Brazil). The tension in each photoelastic model was monitored, photographed and observed using the software Phothoshop 7.0. For qualitative analysis, the area under the implant apex was measured including the green band of the second order fringe of each model using the software Image Tool. After comparison of the areas, the performance generated by each specimen was defined regarding the axial loading. Results: There were alterations in area with different height and thickness of the photoelastic models. It was observed that the group III (30mm in height) presented the smallest area. Conclusion: There was variation in the size of the areas analyzed for different height and thickness of the models and the morphology of the replica may directly influence the result in researches with photoelastic models.

Reconstruction of maxillary ridge atrophy caused by dentoalveolar trauma, using autogenous block bone graft harvested from chin: a case report

Background Dentoalveolar trauma, especially when involving front teeth, negatively affect the patient’s life; in particular, tooth avulsion is a complex injury that affects multiple tissues, and no treatment option offers stable long-term outcomes. The aim of this study was to report a case of reconstruction of atrophic anterior alveolar ridge after tooth loss, performed with autograft harvested from the chin, and subsequent prosthetic rehabilitation with the use of an osseointegrated implant. Case report A 23-years-old Caucasian girl, presented an atrophic alveolar bone in the area of tooth 11, as a result of tooth resorption 10 years after a tooth reimplantation procedure. Reconstruction was performed with autogenous bone harvested from the chin. After 6-months healing period to allow autograft incorporation, a dental implant was inserted. After further 6- months, a screw-retained implant supported metal-ceramic prosthesis was fabricated. Results The prosthetic rehabilitation was successful, and after a follow-up period of 5 years, the achieved result was stable.Conclusion It can be concluded that the autogenous bone graft harvested from the chin, is a safe and effective option for alveolar ridge defects reconstruction, allowing a subsequent placement of a dental implant supporting a prosthetic restoration.

Potential of Bioactive Glass Particles of Different Size Ranges to Affect Bone Formation in Interproximal Periodontal Defects in Dogs

Background: The aim of this study was to compare the potential of bioactive glass particles of different size ranges to affect bone formation in periodontal defects, using the guided tissue regeneration model in dogs. Methods: In six dogs, 2-wall intrabony periodontal defects were surgically created and chronified on the mesial surfaces of mandibular third premolars and first molars bilaterally. After 1 month, each defect was randomly assigned to treatment with bioabsorbable membrane in association with bioactive glass with particle sizes between 300 and 355 mu m (group 1) or between 90 and 710 mu m (group 2), membrane alone (group 3), or negative control (group 4). The dogs were sacrificed 12 weeks after surgeries, and histomorphometric measurements were made of the areas of newly formed bone, new mineralized bone, and bioactive glass particle remnants. Results: With regard to the area of bioactive glass particle remnants, there was a statistically significant difference between groups 1 and 2, favoring group 1. There were greater areas of mineralized bone in groups 1 and 2 compared to groups 3 and 4 (P<0.05). Conclusion: The bioactive glass particles of small size range underwent faster resorption and substitution by new bone than the larger particles, and the use of bioactive glass particles favored the formation of mineralized bone. J Periodontol 2009;80:808-815.

Effect of a low-level laser on bone regeneration after rapid maxillary expansion

Introduction: In this study, we evaluated the effects of a low-level laser on bone regeneration in rapid maxillary expansion procedures. Methods: Twenty-seven children, aged 8 to 12 years, took part in the experiment, with a mean age of 10.2 years, divided into 2 groups: the laser group (n=14), in which rapid maxillary expansion was performed in conjunction with laser use, and the no-laser group (n=13), with rapid maxillary expansion only. The activation protocol of the expansion screw was 1 full turn on the first day and a half turn daily until achieving overcorrection. The laser type used was a laser diode (TWIN Laser; MMOptics, Sao Carlos, Brazil), according to the following protocol: 780 nm wavelength, 40 mW power, and 10 J/cm(2) density at 10 points located around the midpalatal suture. The application stages were 1 (days 1-5 of activation), 2 (at screw locking, on 3 consecutive days), 3, 4, and 5 (7, 14, and 21 days after stage 2). Occlusal radiographs of the maxilla were taken with the aid of an aluminum scale ruler as a densitometry reference at different times: T1 (initial), T2 (day of locking), T3 (3-5 days after T2), T4 (30 days after T3), and T5 (60 days after T4). The radiographs were digitized and submitted to imaging software (Image Tool; UTHSCSA, San Antonio, Tex) to measure the optic density of the previously selected areas. To perform the statistical test, analysis of covariance was used, with the time for the evaluated stage as the covariable. In all tests, a significance level of 5% (P<0.05) was adopted. Results: From the evaluation of bone density, the results showed that the laser improved the opening of the midpalatal suture and accelerated the bone regeneration process. Conclusions: The low-level laser, associated with rapid maxillary expansion, provided efficient opening of the midpalatal suture and influenced the bone regeneration process of the suture, accelerating healing. (Am J Orthod Dentofacial Orthop 2012;141:444-50)