Estudo histomorfométrico do processo de cicatrização óssea em defeitos cirúrgicos de tamanho crítico em calvárias de ratos preenchidos com polímero de mamona

Objectives: The aim of this study was to evaluate the behavior of the polymer histomorphometrically castor during the healing process of defects of critical size calvarial preparations in rats. Materials and Methods: Twenty animals underwent a surgical procedure that was to be held in the calvaria of each animal a critical defect of 8 mm in diameter with a drill trephine. The rats were divided into two groups according to the following procedures: group C received no treatment and the bone defect site was filled with blood clot, group M, the bone defect was filled with castor oil polymer particles. The animals were sacrificed 180 days after the surgical procedures. After routine laboratory procedures the specimens were subjected to analysis histomorphometric. Results: In groups C the newly formed bone tissue was well developed, with adjacent areas of osteoid matrix rich in osteoblasts, and restricted to the vicinity of the edges of the defect. In animals of group M was observed newly formed lamellar bone tissue restricted to the vicinity of the defect edges and particles of polymer Castor distributed throughout the defect. There was a higher percentage of newly formed bone area was statistically significant in group C compared to animals in group M. Conclusions: Within the limits of this study can conclude that the castor oil polymer is biocompatible and had kept the area during the healing of critical size defects in surgically prepared rat calvariae

Regeneração óssea guiada com o uso de matriz óssea bovina desmineralizada em defeitos ósseos experimentais em cães

Background: The principles of tissue regeneration to repair alveolar bone defects are based on the fabrication of a biologic barrier with different biomaterials. Therefore, the present study aimed to investigate the guided bone regeneration (GBR) by using membrane of demineralized bovine bone matrix (DBBM) on experimental defects in tibia of dogs. Methods: Four dogs were used and after anesthesia, shaving and antisepsis, two standardized bone defects were created on the right tibia of each animal with trephine drill. In the Control Group, the defects were filled with blood coagulum, while in the Treated Group, a membrane of DBBM was used to cover the defects. After 90 days, animals were sacrificed. Results: In the Treated Group, 67.4% of new bone formation was observed and, in the Control Group, 32.6% of the bone tissue reabsorbed when compared with initial bone volume. The membrane remained intact and no inflammatory tissue was observed on membrane/ bone interface. Conclusion: It was concluded that the use of DBBM is an osteoconductive material, presents biocompatibility and may be promise option to repair bone defects.

A proporção de plasma rico em plaquetas/enxerto ósseo autógeno pode influenciar o reparo ósseo de defeito de tamanho crítico: estudo radiográfico em calvária de ratos

Introduction: It has been suggested that bone grafts combined with platelet rich plasma (PRP) present greater density than bone grafts without PRP after healing process. However, this result may depend on proportion PRP/autogenous bone (AB) graft used. Objective: The purpose of this study was to analyze radiographically the influence of the proportion PRP/particulate autogenous bone (AB) graft on bone healing in surgically created criticalsize defects (CSD) in rat calvaria. Material and method: 50 rats were divided into 5 groups: C, AB, AB/PRP-50, AB/PRP-100 and AB/PRP-150. A 5 mm diameter CSD was created in the calvarium of each animal. In Group C the defect was filled by blood clot only. In Group AB the defect was filled with 0.01 mL of AB graft. In groups AB/ RP-50, AB/PRP-100 and AB/PRP-150 the defects were filled with 0.01 mL of AB graft combined with 50, 100 and 150 µL of PRP, respectively. All animals were euthanized at 30 days post-operative. Standardized radiographic images of the rat calvaria were obtained using Digora System (Soredex, Finland). The images were evaluated by three examiners using scores that indicated the percentage of radiopacity of the defect. The obtained data were subjected to statistical analysis (Kruskal-Wallis test, p < 0.05). Result: Group C presented radiopacity significantly lower than groups AB, AB/PRP-50, AB/PRP-100 and AB/PRP-150. Group AB/PRP-50 showed radiopacity significantly greater than groups AB/PRP-100 and AB. Conclusion: It can be concluded that the proportion PRP/AB graft influences bone healing in CSD in rat calvaria.

Regeneração óssea guiada em implantodontia: relato de caso

Objective: The use of methods for tissue regeneration has been widely applied in Implantology, in clinical situations with disabilities or anatomical limitations that prevent the placement of osseointegrated dental implants. The evolution of the development of biomaterials revolutionized this therapeutic modality, facilitating the resolution of clinical cases with tissue deficiencies. Thus, this study aimed to describe a clinical case approaching the methods, techniques, and materials used in guided bone regeneration applied to Implantology. Case report: A clinical case of a patient who received a Morse taper dental implant (region 15) is described. The use of biomaterial and membrane on the buccal wall of the socket was required. After the osseointegration period, a reopening surgery was performed, and an immediate provisional implant was produced. After 2 months of follow-up, the final prosthesis was made involving other adjacent elements. Final considerations: The guided bone regeneration technique employed showed satisfactory performance. The patient was positive regarding esthetics and function. However, more controlled studies with longer follow-up period are needed for analyses of predictability

Osteotomia com brocas de alta e baixa rotação, com e sem refrigeração líquida: estudo histopatológico duplo cego em suínos

Nine swines of the White Large race were used for study of the process of bony repair after osteotomy with rotating instruments of low and high-speed, with and without liquid refrigeration. The authors observed, through double blind histopathological analysis, that in every analyzed operative time, the bony repair was processed more appropriately when the osteotomies were accomplished with liquid refrigeration. In the final postoperative times, of fourteen and twenty-eight days, the best results of bony repair were observed in the osteotomy accomplished with drills of low rotation and with liquid refrigeration, while in the postoperative time of seven days, the results of the bony repair of osteotomy accomplished with drills of high rotation and with liquid refrigeration they were better. The authors consider, still, that the process of bony repair in swine display larger proximity in the histological pictures among the studied variables, when compared with other biological models. As most of the works told in the literature it refers the use of dogs, the authors consider that the present work guarantees your importance in the sense of looking for a closer model of the man.

Natural rubber latex coated with calcium phosphate for biomedical application

Natural rubber latex (NRL) is a flexible biomembrane that possesses angiogenic properties and has recently been used for guided bone regeneration, enhancing healing without fibrous tissue, allergies or rejection. Calcium phosphate (Ca/P) ceramics have chemical, biological, and mechanical properties similar to mineral phase of bone, and ability to bond to the host tissue, although it can disperse from where it is applied. Therefore, to create a composite that could enhance the properties of both materials, NRL biomembranes were coated with Ca/P. NRL biomembranes were soaked in 1.5 times concentrated SBF solution for seven days, avoiding the use of high temperatures. SEM showed that Ca/P has been coated in NRL biomembrane, XRD showed low crystallinity and FTIR showed that is the carbonated type B. Furthermore, hemolysis of erythrocytes, quantified spectrophotometrically using materials (Ca/P, NRL, and NRL + Ca/P) showed no hemolytic effects up to 0.125 mg/mL (compounds and mixtures), indicating no detectable disturbance of the red blood cell membranes. The results show that the biomimetic is an appropriate method to coat NRL with Ca/P without using high temperatures, aiming a new biomembrane to improve guided bone regeneration.

Estudo comparativo do desempenho de diferentes membranas oclusivas de Látex Natural no reparo ósseo

Introduction: In dentistry, alveolar bone resorption is a limiting factor in the well being of individuals directly interfering in the stomatognathic system, causing problems in the context of overall health. Aiming to promote biological methods that can stimulate bone regeneration, several biomimetic strategies have been developed by the use of diverse materials possible to the bone matrix, culminating in the development of techniques that promote such repair. Objective: This work is a comparative study of the performance of films made with latex as occlusive membrane for Guided Bone Regeneration (GBR) procedure in three preparations: Latex preserved in ammonia, produced by Latex rubber clones IAN873 and PR255 polymerized immediately after collection and without use ammonia as a preservative. Methods: Sixty Wistar rats were randomly divided into 4 groups of 15 animals in which bone defects of critical size (8mm diameter) were made surgically in the skull. Group A was treated by GBR through the membrane latex preserved with ammonia, Group B received the membrane made of latex from IAN873, Group C, the membrane clone PR255 and group D was not treated by GBR. After a period of 7, 15 and 50 days, 5 animals from each group were euthanized, and specimens containing bone defect collected for microscopic examination (descriptive histology and histomorphometry). Results: The results showed that after 50 days there was bone formation in higher proportions in group D (p <0.05, ANOVA followed by Tukey), suggesting that further experiments should be conducted to conclude about the presence of ammonia and the influence of kind of rubber. Conclusion: GOR is a procedure proven effective in the treatment of bone defects. Therefore, further experiments should be conducted to reach a conclusion regarding the presence of ammonia in the latex composition for the manufacture of membranes, as well as the difference induced by the species of rubber.

Scaffold nanoestruturado utilizando-se celulose bacteriana/fosfatos de cálcio para regeneração óssea

Pós-graduação em Biotecnologia - IQ

Cimentos bioativos injetáveis funcionalizados com peptídeo osteogênico para reparação óssea

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência da inalação da fumaça de cigarro na regeneração óssea guiada em enxertos autógenos onlay em ratas com deficiência estrogênica: estudo histomorfométrico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência da inalação da fumaça de cigarro na regeneração óssea guiada em enxertos autógenos onlay em ratas com deficiência estrogênica: estudo histomorfométrico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Platelet-rich plasma stimulates cytokine expression and alkaline phosphatase activity in osteoblast-derived osteosarcoma cells

Objective: The aim of this study was to investigate the effects of PRP on SAOS-2 cells in terms of cytokine expression, cell activity and oxidative stress. Design: Cell line SAOS-2 (1 x 10(5) cells/mL) were grown in culture medium alpha-MEM with 10% FBS for 24 h and stimulated (or not) with PRP at concentrations of 3, 10 and 20%, LPS (E. coli, 10 g/mL) and IL-1 beta (1 mg/mL) for 24 h. The supernatant was collected and analyzed for the expression of cytokines in a panel array, ALP using a commercial kit and NO2- with Griess reaction method. Also, the cells were analyzed using Western blot for RANKL and slot blotting for nitrotyrosine expression. Result: There were no significant differences amongst the groups in terms of NO2-, protein nitrotyrosine content and RANKL expression. However, all stimuli increased ALP activity and in case of PRP, it was in a dose-dependent manner (p < 0.001). Also, all stimuli induced an increase in cytokines and chemokines expression, but only PRP promoted an increase of component C5, sICAM-1 and RANTES expression. Whilst IL-1 receptor antagonist (IL-1ra) expression was down-regulated by PRP, both LPS and IL-1 beta caused up-regulation of this cytokine. Conclusions: PRP can stimulate osteoblast activity and cytokine/chemokine release, as well as indicate some of the mediators that can (and cannot) be involved in this activation. (C) 2012 Elsevier Ltd. All rights reserved.

Desenvolvimento de um modelo experimental de falha óssea infectada na ulna de coelhos

OBJETIVO: Desenvolver um modelo experimental que permita estudar a regeneração de grandes falhas ósseas em condições de infecção. MÉTODO: Falhas ósseas segmentares de 15mm foram criadas cirurgicamente na ulna de 12 coelhos e inoculadas com 5x10(8) unidades formadoras de colônia (UFC) de S. aureus. O desbridamento da infecção foi realizado duas semanas após, seguida da aplicação sistêmica de gentamicina por quatro semanas. Os animais foram acompanhados por um período de 12 semanas para avaliação do controle da infecção e da regeneração óssea. RESULTADOS: A regeneração espontânea foi inferior a 25% do defeito na avaliação radiográfica e histológica. CONCLUSÃO: A Falha óssea infectada de 15mm na ulna de coelhos é incapaz de alcançar a regeneração completa sem tratamentos adicionais. Nível de Evidência V, Estudo experimental.

New insights in osteogenic differentiation revealed by mass spectrometric assessment of phosphorylated substrates in murine skin mesenchymal cells

Abstract Background Bone fractures and loss represent significant costs for the public health system and often affect the patients quality of life, therefore, understanding the molecular basis for bone regeneration is essential. Cytokines, such as IL-6, IL-10 and TNFα, secreted by inflammatory cells at the lesion site, at the very beginning of the repair process, act as chemotactic factors for mesenchymal stem cells, which proliferate and differentiate into osteoblasts through the autocrine and paracrine action of bone morphogenetic proteins (BMPs), mainly BMP-2. Although it is known that BMP-2 binds to ActRI/BMPR and activates the SMAD 1/5/8 downstream effectors, little is known about the intracellular mechanisms participating in osteoblastic differentiation. We assessed differences in the phosphorylation status of different cellular proteins upon BMP-2 osteogenic induction of isolated murine skin mesenchymal stem cells using Triplex Stable Isotope Dimethyl Labeling coupled with LC/MS. Results From 150 μg of starting material, 2,264 proteins were identified and quantified at five different time points, 235 of which are differentially phosphorylated. Kinase motif analysis showed that several substrates display phosphorylation sites for Casein Kinase, p38, CDK and JNK. Gene ontology analysis showed an increase in biological processes related with signaling and differentiation at early time points after BMP2 induction. Moreover, proteins involved in cytoskeleton rearrangement, Wnt and Ras pathways were found to be differentially phosphorylated during all timepoints studied. Conclusions Taken together, these data, allow new insights on the intracellular substrates which are phosphorylated early on during differentiation to BMP2-driven osteoblastic differentiation of skin-derived mesenchymal stem cells.

Reconstructive Microsurgery and Tissue Engineering in Musculo-Skeletal Oncology - Innovative Techniques

Tumors involving bone and soft tissues are extremely challenging situations. With the recent advances of multi-modal treatment, not only the type of surgery has moved from amputation to limb-sparing procedures, but also the survivorship has improved considerably and reconstructive techniques have the goal to allow a considerably higher quality of life. In bone reconstruction, tissue engineering strategies are the main area of research. Re-vascularization and re-vitalisation of a massive allograft would considerably improve the outcome of biological reconstructions. Using a rabbit animal model, in this study we showed that, by implanting a vascular pedicle inside a weight bearing massive cortical allograft, the bone regeneration inside the allograft was higher compared to the non-vascularized implants, given the patency of the vascular pedicle. Improvement in the animal model and the addition of Stem Cells and Growth factors will allow a further improvement in the results. In soft tissue tumors, free and pedicled flaps have been proven to be of great help as reconstruction strategies. In this study we analyzed the functional and overall outcome of 14 patients who received a re-innervated vascularized flap. We have demonstrated that the use of the innovative technique of motor re-innervated muscular flaps is effective when the resection involves important functional compartments of the upper or lower limb, with no increase of post-operative complications. Although there was no direct comparison between this type of reconstruction and the standard non-innervated reconstruction, we underlined the remarkable high overall functional scores and patient satisfaction following this procedure.