Immunolocalization of markers for bone formation during guided bone regeneration in osteopenic rats

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

Bacterial cellulose biocomposites for guided tissue regeneration

Bacterial cellulose (BC) has become established as a remarkably versatile biomaterial and can be used in a wide variety of scientific applications, especially for medical devices. In this work, the bacterial cellulose fermentation process is modified by the addition of chondroitin sulfate and hyaluronic acid (1% w/w) to the culture medium before the bacteria is inoculated. Besides, biomimetic precipitation of calcium phosphate of biological interest from simulated body fluid on bacterial cellulose was studied. Chondroitin sulfate and hyaluronic acid influences in bacterial cellulose were analyzed using transmission infrared spectroscopy (FTIR), XRD (X-ray diffraction) and scanning electron microscopy (SEM). FTIR analysis showed interaction between bacterial cellulose nanobiocomposites and calcium phosphate and XRD demonstrated amorphous calcium phosphate and calcium chloride on bacterial cellulose nanobiocomposites. SEM images confirmed incorporation of calcium phosphate in bacterial cellulose nanobiocomposites surface with different calcium phosphate particles morphology.

Skeletal muscle degeneration and regeneration after injection of bothropstoxin-II, a phospholipase A2 isolated from the venom of the snake Bothrops jararacussu

A myotoxic phospholipase A2, named bothropstoxin II (BthTX-II), was isolated from the venom of the South American snake Bothrops jararacussu and the pathogenesis of myonecrosis induced by this toxin was studied in mice. BthTX-II induced a rapid increase in plasma creatine kinase levels. Histological and ultrastructural observations demonstrate that this toxin affects muscle fibers by first disrupting the integrity of plasma membrane, as delta lesions were the earliest morphological alteration and since the plasma membrane was interrupted or absent in many portions. In agreement with this hypothesis, BthTX-II released peroxidase entrapped in negatively charged multilamellar liposomes and behaved as an amphiphilic protein in charge shift electrophoresis, an indication that its mechanism of action might be based on the interaction and disorganization of plasma membrane phospholipids. Membrane damage was followed by a complex series of morphological alterations in intracellular structures, most of which are probably related to an increase in cytosolic calcium levels. Myofilaments became hypercontracted into dense clumps which alternated with cellular spaces devoid of myofibrillar material. Later on, myofilaments changed to a hyaline appearance with a more uniform distribution. Mitochondria were drastically affected, showing high amplitude swelling, vesiculation of cristae, formation of flocculent densities, and membrane disruption. By 24 hr, abundant polymorphonuclear leucocytes and macrophages were observed in the interstitial space as well as inside necrotic fibers. Muscle regeneration proceeded normally, as abundant myotubes and regenerating myofibers were observed 7 days after BthTX-II injection. By 28 days regenerating fibers had a diameter similar to that of adult muscle fibers, although they presented two distinctive features: central location of nuclei and some fiber splitting. This good regenerative response may be explained by the observation that BthTX-II does not affect blood vessels, nerves, or basal laminae. © 1991.

Preparation and characterization of a bacterial cellulose/silk fibroin sponge scaffold for tissue regeneration

Bacterial cellulose (BC) and silk fibroin (SF) are natural biopolymers successfully applied in tissue engineering and biomedical fields. In this work nanocomposites based on BC and SF were prepared and characterized by scanning electron microscopy (SEM), infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and thermogravimetric analysis (TGA). In addition, the investigation of cytocompatibility was done by MTT, XTT and Trypan Blue dye technique. Cellular adhesion and proliferation were detected additionally. The evaluation of genotoxicity was realized by micronucleus assay. In vitro tests showed that the material is non-cytotoxic or genotoxic. SEM images revealed a greater number of cells attached at the BC/SF:50% scaffold surface than the pure BC one, suggesting that the presence of fibroin improved cell attachment. This could be related to the SF amino acid sequence that acts as cell receptors facilitating cell adhesion and growth. Consequently, BC/SF:50% scaffolds configured an excellent option in bioengineering depicting its potential for tissue regeneration and cultivation of cells on nanocomposites.

Bone regeneration in surgically created deffects filled with autogenous bone: epifluorescence microscopy analysis in rats

Although the search for the ideal bone substitute has been the focus of a large number of studies, autogenous bone is still the gold standard for the filling of defects caused by pathologies and traumas, and mainly, for alveolar ridge reconstruction, allowing the titanium implants installation. OBJECTIVES: The aim of this study was to evaluate the dynamics of autogenous bone graft incorporation process to surgically created defects in rat calvaria, using epifluorescence microscopy. MATERIAL AND METHODS: Five adult male rats weighing 200-300 g were used. The animals received two 5-mm-diameter bone defects bilaterally in each parietal bone with a trephine bur under general anesthesia. Two groups of defects were formed: a control group (n=5), in which the defects were filled with blood clot, and a graft group (n=5), in which the defects were filled with autogenous bone block, removed from the contralateral defect. The fluorochromes calcein and alizarin were applied at the 7th and 30th postoperative days, respectively. The animals were killed at 35 days. RESULTS: The mineralization process was more intense in the graft group (32.09%) and occurred mainly between 7 and 30 days, the period labeled by calcein (24.66%). CONCLUSIONS: The fluorochromes showed to be appropriate to label mineralization areas. The interfacial areas between fluorochrome labels are important sources of information about the bone regeneration dynamics.

Combined therapy with mineral trioxide aggregate, and guided tissue regeneration for a large radicular cyst: a 13-year follow-u

Biomaterials such as membrane barriers and/or bone grafts are often used to enhance periapical new bone formation. A combination of apical surgery and these biomaterials is one of the latest treatment options for avoiding tooth extraction. In case of periapical lesions, guided tissue regeneration (GTR) is attempted to improve the self-regenerative healing process by excluding undesired proliferation of the gingival connective tissue or migration of the oral epithelial cells into osseous defects. In many cases, GTR is necessary for achieving periodontal tissue healing. This report describes the healing process after surgery in a challenging case with a long-term followup. In this case report, endodontic surgery was followed by retrograde sealing with mineral trioxide aggregate (MTA) in the maxillary right central incisor and left lateral incisor. Apicectomy was performed in the maxillary left central incisor and a 1-mm filling was removed. The bone defect was filled with an anorganic bone graft and covered with a decalcified cortical osseous membrane. No intraoperative or postoperative complications were observed. After 13 years of follow-up, the patient showed no clinical signs or symptoms associated with the lesion and radiographic examination showed progressive resolution of radiolucency. In conclusion, the combination of apical surgery and regenerative techniques can successfully help the treatment of periapical lesions of endodontic origin and is suitable for the management of challenging cases

Root reconstructed with mineral trioxide aggregate and guided tissue regeneration in apical surgery: a 5-year follow-up

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

Ecological optimization of an irreversible Brayton cycle with regeneration, inter-cooling and reheating

A mathematical model is developed for an irreversible Brayton cycle with regeneration, inter-cooling and reheating. The irreversibility are from the thermal resistance in the heat exchangers, the pressure drops in pipes, the non-isentropic behavior in the adiabatic expansions and compressions and the heat leakage to the cold source. The cycle is optimized by maximizing the ecological function, which is achieved by the search for optimal values for the temperatures of the cycle and for the pressure ratios of the first stage compression and the first stage expansion. The advantages of using the regenerator, intercooler and reheater are presented by comparison with cycles that do not incorporate one or more of these processes. Optimization results are compared with those obtained by maximizing the power output and it is concluded that the point of maximum ecological function has major advantages with respect to the entropy generation rate and the thermal efficiency, at the cost of a small loss in power.