Orthodontic force increases interleukin-1β and tumor necrosis factor-α expression and alveolar bone loss in periodontitis

Background: The present study aims to evaluate the effects of orthodontic movement (OM) on the periodontal tissues of rats with ligature-induced periodontal disease. Methods: Eighty-eight rats were divided into four groups: 1) negative control (sham operated); 2) periodontal disease; 3) OM; and 4) periodontal disease followed by OM (OMP). Rats were sacrificed 3 hours or 1, 3, or 7 days after OM commencement. Bone volume fraction (BVF) and bone mineral density (BMD) were assessed in hemimaxillae by microcomputed tomography analysis. Expression of the proinflammatory cytokines interleukin (IL)-1β and tumor necrosis factor (TNF)-α were evaluated in gingival samples by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, and in the furcation region by immunohistochemistry analysis (IHC). Results: The OMP group had lower BVF and BMD levels compared to the other groups at day 7 (P <0.05). Maximum messenger ribonucleic acid expression of both cytokines was observed in the OMP group at day 1 (P <0.05). In the same period, all proteins were expressed in high levels for all test groups compared to the control group. The number of cells positive for IL-1β and TNF-α by IHC was highest in the OMP group at day 1, with progressive reduction thereafter. Conclusion: The results suggest that OM acts synergistically with periodontal disease in periodontal breakdown through upregulation of proinflammatory cytokines.

Histologic Characterization of Engineered Tissues in the Canal Space of Closed-apex Teeth with Apical Periodontitis

Introduction: The aim of this study was to investigate the capacity of endodontic regenerative procedures combining an induced blood clot, platelet-rich plasma (PRP), and bone marrow aspirate (BMA) to regenerate dental pulp in canine closed-apex necrotic teeth. Methods: Apical periodontitis was induced in 20 upper and lower premolars of 2 dogs. After biomechanical preparation, enlargement to a #60 file, and disinfection with a triantibiotic paste for 28 days, the roots were randomly assigned to 4 treatment groups: blood clot (BC), BC + PRP gel, BC + BMA gel, and BC + BMA/PRP gel. Negative controls were also included. After a 3-month follow-up period, the animals were killed. Results: Histologic analysis showed the presence of newly formed vital tissues (connective, cement-like, and bone-like tissue) in 23 of the 32 treated roots (71.87%). There was no statistically significant difference between the treatment groups. Conclusions: New vital tissues were formed and characterized as connective, cementum-like, or bone-like, but not as pulp-like tissue; PRP and/or BMA did not improve the tissue ingrowth. © 2013 American Association of Endodontists.

Stress analysis in bone tissue around single implants with different diameters and veneering materials: A 3-D finite element study

The aimof this study was to evaluate the stress distribution on bone tissue with a single prosthesis supported by implants of large and conventional diameter and presenting different veneering materials using the 3-D finite elementmethod. Sixteenmodels were fabricated to reproduce a bone block with implants, using two diameters (3.75 × 10 mmand 5.00 × 10 mm), four different veneering materials (composite resin, acrylic resin, porcelain, and NiCr crown), and two loads (axial (200 N) and oblique (100 N)). For data analysis, the maximum principal stress and vonMises criterion were used. For the axial load, the cortical bone in allmodels did not exhibit significant differences, and the trabecular bone presented higher tensile stresswith reduced implant diameter. For the oblique load, the cortical bone presented a significant increase in tensile stress on the same side as the loading for smaller implant diameters. The trabecular bone showed a similar but more discreet trend. There was no difference in bone tissue with different veneering materials. The veneering material did not influence the stress distribution in the supporting tissues of single implant-supported prostheses. The large-diameter implants improved the transference of occlusal loads to bone tissue and decreased stress mainly under oblique loads.Oblique loading was more detrimental to distribution stresses than axial loading. © 2013 Elsevier B.V. All rights reserved.

Comparison of changes in dental and bone radiographic densities in the presence of different soft-tissue simulators using pixel intensity and digital subtraction analyses

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

Oral squamous carcinoma cells secrete RANKL directly supporting osteolytic bone loss

Objective: Local invasion of bone is a frequent complication of oral squamous cell carcinoma (OSCC). Development of these osteolytic lesions is mediated by osteoclasts. Receptor activation of NF-kappa B ligand (RANKL) signaling, counteracted by osteoprotegerin (OPG), regulates osteoclastogenesis. Previous studies in rodent models have demonstrated that inhibition of RANKL decreases tumor growth and lesions within bone. However, the contributory role of OSCC cells to this disease process has yet to be defined.Methods: RANKL expression was assessed in a panel of OSCC cell lines by qPCR, flow cytometry, and ELISA. Induction of osteoclastogenesis was assessed by co-culture with macrophages or with OSCC-derived conditioned medium. In an animal model of bone invasion, nude mice were injected intratibially with UMSCC-11B cells expressing a RANKL luciferase promoter to detect tumor-derived RANKL activity. Osteolytic lesions were analyzed by X-ray, micro-CT, and histological methods. RANKL expression was assessed in human OSCC tissues by immunohistochemistry.Results: We demonstrated that OSCCs express varied levels of all RANKL isoforms, both membrane-bound and soluble RANKL. Both co-culture and treatment with OSCC-conditioned media induced osteoclastogenesis. In mice, we demonstrated human RANKL promoter activity during bone invasion. Over the course of the experiment, animals suffered osteolytic lesions as RANKL-driven luciferase expression increased with time. After 8 weeks, human-derived RANKL was detected in areas of bone resorption by immunohistochemistry. Similar epithelial RANKL expression was detected in human OSCC tissues.Conclusion: These data demonstrate the ability of OSCCs to produce RANKL, directly altering the tumor microenvironment to increase osteoclastogenesis and mediate local bone invasion. (C) 2012 Elsevier Ltd. All rights reserved.

Bone-implant contact around crestal and subcrestal dental implants submitted to immediate and conventional loading

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

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.

Remodeling of cortical and corticocancellous fresh-frozen allogeneic block bone grafts - a radiographic and histomorphometric comparison to autologous bone grafts

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

Anti-inflammatory effect of MAPK phosphatase-1 local gene transfer in inflammatory bone loss

Alveolar bone loss associated with periodontal diseases is the result of osteoclastogenesis induced by bacterial pathogens. The mitogen-activated protein kinase (MAPK) phosphatase 1 (MKP-1) is a critical negative regulator of immune response as a key phosphatase capable of dephosphorylating activated MAPKs. In this study, rat macrophages transduced with recombinant adenovirus (Ad.)MKP-1 specifically dephosphorylated activated MAPKs induced by lipopolysaccharide (LPS) compared with control cells. Bone marrow macrophages from MKP-1 knockout (KO) mice exhibited higher interleukin (IL)-6, IL-10, tumor necrosis factor (TNF)-α, and select chemokine compared with wild-type (WT) mice when stimulated by LPS. In addition, bone marrow cultures from MKP-1 KO mice exhibited significantly more osteoclastogenesis induced by LPS than when compared with WT mice. Importantly, MKP-1 gene transfer in bone marrow cells of MKP-1 KO mice significantly decreased IL-6, IL-10, TNF-α and chemokine levels, and formed fewer osteoclasts induced by LPS than compared with control group of cells. Furthermore, MKP-1 gene transfer in an experimental periodontal disease model attenuated bone resorption induced by LPS. Histological analysis confirmed that periodontal tissues transduced with Ad. MKP-1 exhibited less infiltrated inflammatory cells, less osteoclasts and less IL-6 than compared with rats of control groups. These studies indicate that MKP-1 is a key therapeutic target to control of inflammation-induced bone loss.

Silencing mitogen-activated protein kinase-activated protein kinase-2 arrests inflammatory bone loss

p38 mitogen-activated protein kinases (MAPKs) are critical for innate immune signaling and subsequent cytokine expression in periodontal inflammation and bone destruction. In fact, previous studies show that systemic p38 MAPK inhibitors block periodontal disease progression. However, development of p38 MAPK inhibitors with favorable toxicological profiles is difficult. Here, we report our findings regarding the contribution of the downstream p38 MAPK substrate, mitogen-activated protein kinase-activated protein kinase 2 (MK2 or MAPKAPK-2), in immune response modulation in an experimental model of pathogen-derived lipopolysaccharide (LPS)-induced periodontal bone loss. To determine whether small interfering RNA (siRNA) technology has intraoral applications, we initially validated MK2 siRNA specificity. Then, gingival tissue surrounding maxillary molars of rats was injected with MK2 siRNA or scrambled siRNA at the palatal regions of bone loss. Intraoral tissues treated with MK2 siRNA had significantly less MK2 mRNA expression compared with scrambled siRNA-treated tissues. MK2 siRNA delivery arrested LPS-induced inflammatory bone loss, decreased inflammatory infiltrate, and decreased osteoclastogenesis. This proof-of-concept study suggests a novel target using an intraoral RNA interference strategy to control periodontal inflammation.

Feasibility and safety of intrathecal transplantation of autologous bone marrow mesenchymal stem cells in horses

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

Dimensional changes in soft tissues around dental implants following free gingival grafting: an experimental study in dogs

ObjectiveTo study the buccal dimensional tissue changes at oral implants following free gingival grafting, with or without including the keratin layer, performed at the time of implant installation into alveolar mucosa.Material and methodsThe mandibular premolars and first molars were extracted bilaterally in six Beagle dogs. In the right side of the mandible (Test), flaps were first elevated, and the buccal as well as part of the lingual masticatory mucosa was removed. An incision of the periosteum at the buccal aspect was performed to allow the flap to be coronally repositioned. Primary wound closure was obtained. In the left side, the masticatory (keratinized) mucosa was left in situ, and no sutures were applied (Control). After 3months of healing, absence of keratinized mucosa was confirmed at the test sites. Two recipient sites were prepared at each side of the mandible in the region of the third and fourth premolars. All implants were installed with the shoulder placed flush with the buccal alveolar bony crest, and abutments were connected to allow a non-submerged healing. Two free gingival mucosal grafts were harvested from the buccal region of the maxillary canines. One graft was left intact (gingival mucosal graft), while for the second, the epithelial layer was removed (gingival connective tissue graft). Subsequently, the grafts were fixed around the test implants in position of the third and fourth premolars, respectively. After 3months, the animals were euthanized and ground sections obtained.ResultsSimilar bony crest resorption and coronal extension of osseointegration were found at test and control sites. Moreover, similar dimensions of the peri-implant soft tissues were obtained at test and control sites.ConclusionsThe increase in the alveolar mucosal thickness by means of a gingival graft affected the peri-implant marginal bone resorption and soft tissue recession around implants. This resulted in outcomes that were similar to those at implants surrounded by masticatory mucosa, indicating that gingival grafting in the absence of keratinized mucosa around implants may reduce the resorption of the marginal crest and soft tissue recession.

Surgical Treatment of Severe Frontal Bone Fracture

Craniofacial trauma can lead to several complications. The combined fractures of anterior and posterior walls of the frontal bone are almost always followed by lesions in nasofrontal orifices and disruption of nasofrontal ostia or ducts, a significant factor for the development of early and late complications after sinus fractures. This article reports a case of trauma patient, who underwent neurological evaluation and at first showed good general condition. Computed tomography noted fracture of the anterior and posterior walls of the frontal sinus and small foci of pneumocephalus in the cerebral cortex. The patient was monitored periodically and 9 days after trauma showed increased areas of pneumocephalus in prefrontal cortex, cerebrospinal fluid draining, and large dura mater lesion, with signs of necrosis and inflammation (meningitis). The necrotic tissues were removed, and dura mater was repaired through the approximation with resorbable wire polyglactin 910 5-0, oxidized cellulose application, and bonding with human fibrin sealant (fibrinogen, thrombin, and calcium chloride). Sinusectomy, frontal sinus, and nasofrontal duct obliteration with pedicled pericranium flap were performed. Tomographically, a reanatomization was noted in frontal region, and a 12-month follow-up showed no complication. The use of fibrin glue to repair dura mater lacerations, as well as the pedicle pericranium flap for frontal sinus and nasofrontal duct obliteration, is an efficient method for treating fractures of the frontal bone.

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.

Reconstruction of maxillary ridge atrophy caused by sequela of dentoalveolar trauma with autogenous bone graft harvested from mentum

Dentoalveolar traumatisms, particularly those that affect the anterior teeth, interfere adversely in the patient s life.Among them, tooth avulsion is pointed out because it is characterized as a complex injury that affects multiple tissues, andbecause there is no effective treatment available for its resolution with a stable long-term outcome.Aim/Hypothesis: The aim of the present study was to relate a clinical case of complete reconstruction of atrophy of the alveolarbone corresponding to tooth 11, lost by tooth resorption 10 years after the tooth reimplantation procedure.Material and methods: Reconstruction was performed with autogenous bone harvested from the mentum donor site. Surgicalaccess began in the receptor area with a Newman mucoperiosteal incision using a scalpel blade 15 mounted in a scalpel handlefor detachment and exposure of the receptor site. Extensive bone resorption was observed in the vestibular-palatine direction,proved by the thinness of the receptor bed. Decorticalization of the vestibular bone plate was performed. After preparing thereceptor bed, and incision was made in the mucosa in the depth of the anterior vestibular fornix, then a perpendicular muscleperiostealincision to detach and exposure the donor area. The bone graft necessary for reconstruction of the donor area wasdelimited, followed by monocortical osteotomy and the monocortical graft was removed. The next stage was to perform shapingfor passive graft accommodation and fixation by means of two bicortical screws. After fixation of the graft the sharp angles wererounded off in order to avoid possible exposure and/or fenestrations of the reconstructed area, then the receptor and donor areawere sutured. After the 6-month period to allow incorporation of the autogenous graft, an osseointegrated dental implant wasinserted. At the end of the 6-month period of waiting for osseointegration to occur, the process of fabricating the screw-retainedmetal ceramic