Estudo da descelularização tecidual na produção de arcabouços biológicos para enxertos

The regeneration of bone defects with loss of substance remains as a therapeutic challenge in the medical field. There are basically four types of grafts: autologous, allogenic, xenogenic and isogenic. It is a consensus that autologous bone is the most suitable material for this purpose, but there are limitations to its use, especially the insufficient amount in the donor. Surveys show that the components of the extracellular matrix (ECM) are generally conserved between different species and are well tolerated even in xenogenic recipient. Thus, several studies have been conducted in the search for a replacement for autogenous bone scaffold using the technique of decellularization. To obtain these scaffolds, tissue must undergo a process of cell removal that causes minimal adverse effects on the composition, biological activity and mechanical integrity of the remaining extracellular matrix. There is not, however, a conformity among researchers about the best protocol for decellularization, since each of these treatments interfere differently in biochemical composition, ultrastructure and mechanical properties of the extracellular matrix, affecting the type of immune response to the material. Further down the arsenal of research involving decellularization bone tissue represents another obstacle to the arrival of a consensus protocol. The present study aimed to evaluate the influence of decellularization methods in the production of biological scaffolds from skeletal organs of mice, for their use for grafting. This was a laboratory study, sequenced in two distinct stages. In the first phase 12 mice hemi-calvariae were evaluated, divided into three groups (n = 4) and submitted to three different decellularization protocols (SDS [group I], trypsin [Group II], Triton X-100 [Group III]). We tried to identify the one that promotes most efficient cell removal, simultaneously to the best structural preservation of the bone extracellular matrix. Therefore, we performed quantitative analysis of the number of remaining cells and descriptive analysis of the scaffolds, made possible by microscopy. In the second stage, a study was conducted to evaluate the in vitro adhesion of mice bone marrow mesenchymal cells, cultured on these scaffolds, previously decellularized. Through manual counting of cells on scaffolds there was a complete cell removal in Group II, Group I showed a practically complete cell removal, and Group III displayed cell remains. The findings allowed us to observe a significant difference only between Groups II and III (p = 0.042). Better maintenance of the collagen structure was obtained with Triton X-100, whereas the decellularization with Trypsin was responsible for the major structural changes in the scaffolds. After culture, the adhesion of mesenchymal cells was only observed in specimens deccelularized with Trypsin. Due to the potential for total removal of cells and the ability to allow adherence of these, the protocol based on the use of Trypsin (Group II) was considered the most suitable for use in future experiments involving bone grafting decellularized scaffolds

Estudo da detecção preventiva da osteoporose pela aplicação da variação da permissividade elétrica a partir da sondagem eletromagnética

This paper proposes a method based on the theory of electromagnetic waves reflected to evaluate the behavior of these waves and the level of attenuation caused in bone tissue. For this, it was proposed the construction of two antennas in microstrip structure with resonance frequency at 2.44 GHz The problem becomes relevant because of the diseases osteometabolic reach a large portion of the population, men and women. With this method, the signal is classified into two groups: tissue mass with bony tissues with normal or low bone mass. For this, techniques of feature extraction (Wavelet Transform) and pattern recognition (KNN and ANN) were used. The tests were performed on bovine bone and tissue with chemicals, the methodology and results are described in the work

Medição da densidade óssea através de ondas eletromagnéticas e classificação por MVS

This work holds the purpose of presenting an auxiliary way of bone density measurement through the attenuation of electromagnetic waves. In order to do so, an arrangement of two microstrip antennas with rectangular configuration has been used, operating in a frequency of 2,49 GHz, and fed by a microstrip line on a substrate of fiberglass with permissiveness of 4.4 and height of 0,9 cm. Simulations were done with silica, bone meal, silica and gypsum blocks samples to prove the variation on the attenuation level of different combinations. Because of their good reproduction of the human beings anomaly aspects, samples of bovine bone were used. They were subjected to weighing, measurement and microwave radiation. The samples had their masses altered after mischaracterization and the process was repeated. The obtained data were inserted in a neural network and its training was proceeded with the best results gathered by correct classification on 100% of the samples. It comes to the conclusion that through only one non-ionizing wave in the 2,49 GHz zone it is possible to evaluate the attenuation level in the bone tissue, and that with the appliance of neural network fed with obtained characteristics in the experiment it is possible to classify a sample as having low or high bone density

Atividade biológica de células-tronco da polpa de dentes decíduos humanos submetidas à criopreservação

Dental pulp stem cells have been widely investigated because of their ability to differentiate into both dental and non-dental cells, with potential use in therapies involving tissue engineering. The technique of cell cryopreservation represents a viable alternative for the conservation of these cells, since it stops reversibly, in a controlled manner, all of cell biological functions in an ultra low temperature. The present study aimed to evaluate, using in vitro experiments, the influence of a cryopreservation protocol on the biologic acti vity of stem cells from human exfoliated deciduous teeth (SHED). Cells obtained from the pulp of three deciduous teeth on end-stage exfoliation or with indicated extraction were expanded in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. At second subculture (P2), a group of cells were submitted to cryopreservation for 30 days in 10% DMSO diluted in fetal bovine serum, at -80º C, while the remind cells continued under normal conditions of cell culture. Cell proliferation was evaluated in both groups (not cryopreserved or cryopreserved) by Trypan blue stain essay at intervals of 24, 48 and 72h after plating. Cell cycle analysis of SHEDs submitted or not to the cryopreservation protocol was performed in the same intervals. Events related to cell death were studied by Annexyn V and PI expression under flow cytometry at the intervals of 24 and 72h. The presence of nuclear morphological changes was evaluated by DAPI staining at 72h interval. It was observed that both groups exhibited an upward cell proliferation curve, without considerable changes in cell viability throughout the experiment. The distribution of cell in the cell cycle phasis was consistent with cell proliferation in both groups. There were no nuclear morphological damages in the end range of the experiment. therefore, it is concluded that the proposed cryopreservation protocol is efficient for storing the studied cell type, allowing its use in future experimental studies