RESUMO: O objetivo deste estudo foi avaliar o efeito das células osteoprogenitoras derivadas de células tronco mesenquimais do tecido adiposo (CO-CTM-AD) no tratamento de defeitos ósseos críticos de cães. As células tronco mesenquimais do tecido adiposo (CTM-AD) foram submetidas à diferenciação osteogênica por 21 dias e usadas no tratamento de defeitos ósseos em rádios de cães. Foram constituídos três grupos experimentais: defeitos ósseos tratados com CO-CTM-AD (OC), defeitos preenchidos com osso autógeno (C+) e defeitos não preenchidos (C-). A regeneração óssea foi avaliada por meio de exames radiográficos, densitométricos e histomorfométricos. A área de neoformação óssea foi maior no grupo OC em relação ao grupo C- no 15o dia de pós-operatório. Os defeitos tratados com CO-CTM-AD mostraram maior neovascularização que os demais grupos aos 90 dias de avaliação. Conclui-se que o tratamento com CO-CTM-AD aumentou a área de osso neoformado no 15o dia de pós-operatório, mas não foi suficiente para que houvesse a completa união óssea em defeitos ósseos críticos no rádio de cães aos 90 dias.

ABSTRACT: The aim of this study was to evaluate the effect of osteoprogenitor cells derived from mesenchymal stem cells from adipose tissue (OC-AD-MSCs), and differentiated into osteoblasts, in the treatment of critical bone defects in dogs. Adipose tissue derived mesenchymal stem cells (AD-MSCs) were subjected to osteogenic differentiation for 21 days and used in the treatment of bone defects in dogs radius. Either three experimental groups were bone defects treated with OC-AD-MSCs (OC), defects filled with autogenous bone (Control- C +), or empty defects (Control- C -). Bone regeneration was assessed by radiology, densitometry, and histomorphometry. The area of new bone formation was higher in the OC group compared to the control group (C-) on postoperative day 15. Defects treated with OC-AD-MSCs showed greater neovascularization than the other two groups at 90 days. We concluded that treatment with OC-AD-MSCs increased the area of new bone formation 15 days after surgery; however, it didn’t complete the bone union in critical bone defects in the radius of dogs at 90 days.

The aim of this work was the partial purification and subsequent evaluation of chitinase expression during the various growth phases of Paracoccidioides brasiliensis. Initially, PbCTS1r was expressed as a recombinant protein and displayed enzymatic activity against 4-MU-[N-acetylglucosamine (GlcNAc)]3 and 4-MU-(GlcNAc)2. Two proteins, 45 kDa and 39 kDa in size, were partially purified from P. brasiliensis yeast crude extract using cation-exchange chromatography coupled with HPLC and were characterised as PbCTS1 and PbCTS2, respectively. Anti-PbCTS1r antibody recognised two proteins in the crude extracts of yeast and the transitional stage between mycelial and yeast phases. In crude extracts of mycelium, only the 45 kDa protein was detected. However, quantitative real-time polymerase chain reaction led to the detection of small quantities of Pbcts2 transcript in the mycelial phase. In the yeast cell wall extract, only the 39 kDa protein was detected. Moreover, both proteins were secreted by the yeast parasitic phase, suggesting that these proteins participate in the modulation of the fungal environment. Phylogenetic analysis of the predicted PbCTS1 and PbCTS2 proteins indicated that they code for distinct chitinases in P. brasiliensis. During evolution, P. brasiliensis could have acquired the paralogues Pbcts1 and Pbcts2 for growth and survival in diverse environments in both saprophytic and parasitic phases.

A porous biodegradable polyurethane nanocomposite based on poly(caprolactone) (PCL) and nanocomponents derived from montmorillonite (Cloisite®30B) was synthesized and tested to produce information regarding its potential use as a scaffold for tissue engineering. Structural and morphological characteristics of this nanocomposite were studied by infrared spectroscopy (FTIR), X-ray diffraction (XRD), small angle X-ray scattering (SAXS) and scanning electron microscopy (SEM). The reaction between polyurethane oligomers with isocyanate endcapped chains and water led to the evolution of CO2, which was responsible for building interconnected pores with sizes ranging from 184 to 387 μm. An in vitro cell-nanocomposite interaction study was carried out using neonatal rat calvarial osteoblasts. The ability of cells to proliferate and produce an extracellular matrix in contact with the synthesized material was assessed by an MTT assay, a collagen synthesis analysis, and the expression of alkaline phosphatase. In vivo experiments were performed by subcutaneously implanting samples in the dorsum of rats. The implants were removed after 14, 21, and 29 days, and were analyzed by SEM and optical microscopy after tissue processing. Histology crosssections and SEM analyses showed that the cells were able to penetrate into the material and to attach to many location throughout the pore structure. In vitro and in vivo tests demonstrated the feasibility for polyurethane nanocomposites to be used as artificial extracellular matrices onto which cells can attach, grow, and form new tissues.

Células do baço de camundongos foram examinadas aos 5, 10, 20 e 25 dias pós-infecção (dpi) com Dermatobia hominis e examinadas aos 5, 10, 15, 30 e 60 dias pós-emergência da larva (dpel). As células foram cultivadas em meio RPMI-1640 contendo, ou não (controle), produtos de secreção das larvas (PSL) de D. hominis com idade de 5, 10, 15, 20 e 25 dias. Em cada grupo com cinco camundongos testados nos meios de cultura, o número de células foi significativo para 25 dpi, com crescente aumento na seguinte ordem: PSL-10 d, -25 d, -5 d, -20 d, -15 d e RPMI. Resultados significantes foram também observados nos testes entre cada meio contendo células tanto de camundongos dpi ou dpel. Em cada dpel grupo versus meio significância foi somente verificada para 10 dpel, na ordem crescente: PSL-5 d, -20 d, -25 d, -10 d, -15 d e RPMI. Testes comparativos foram também realizados entre grupos. O PSL foi analisado sob SDS-PAGE. Os resultados provam que a miíase causou depleção de células do baço, particularmente sob efeito do PSL-10 e -15, mas ocorreu normalidade do número de células aos 60 dpel. Este ensaio in vitro pode representar uma resposta imune sistêmica na relação PSL-D. hominis-hospedeiro.

Spleen cells from mice were examined at 5, 10, 15, 20 and 25 days post-infection (dpi) with Dermatobia hominis larva and at 5, 10, 15, 30 and 60 days post-larval emergence (dple). Cell proliferation in vitro assays were carried out with RPMI-1640 medium and larval secretory product (LSP) of D. hominis at 5, 10, 15, 20 and 25 days. When each group of mice was tested against each medium, significance was only seen for 25 dpi, with increasing order: LSP-10 d, -25 d, -5 d, -20 d, -15 d and RPMI. Significant results were also observed when each medium was tested against mice at each dpi or dple. Each dple group vs. each medium produced significant results only for 10 dple, with increasing order: LSP-5 d, -20 d, -25 d, -10 d, -15 d and RPMI. Comparative tests were also carried out between groups to refine certain observations. The LSPs were also analyzed using SDS-PAGE. The results prove that myiasis caused depletion of spleen cells, particularly under the effect of the LSP-10 and -15, but the cells tended to increase up to 60 dple. This in vitro assay may represent the real systemic immune response in the relationship LSP-D. hominis-host.

Symptomatic prostatic paracoccidioidomycosis (PCM) is a very rare condition; however, it may express as a typical benign prostatic hyperplasia or a simulating prostatic adenocarcinoma. This case report presents PCM mimicking prostatic adenocarcinoma. The purpose of this paper is to call the general physician's attention to this important differential diagnosis.

Células-tronco têm sido estudadas em várias áreas da Medicina e suas aplicações brevemente deverão estar incorporadas à prática clínica. O dano retiniano pela morte neuronal é considerado incurável devido a pobre capacidade regenerativa do sistema nervoso central. A capacidade das células-tronco em regenerar tecidos, assim como sua plasticidade, faz que estas sejam uma fonte potencial de células para a regeneração retiniana. Células-tronco são muito promissoras para o tratamento das distrofias retinianas, como a retinose pigmentar e outras doenças neurodegenerativas, que podem evoluir para cegueira. As células-tronco mesenquimais são o tipo mais provável de células-tronco a serem utilizadas na prática clínica devido a sua fácil acessibilidade e multipotencialidade de diferenciação em vários tecidos. As células-tronco mesenquimais são células clonogênicas, não-hematopoiéticas, localizadas na medula óssea. Desde que seja proporcionado um microambiente apropriado, estas células podem se diferenciar em cardiomiócitos e até mesmo em células de origem não-mesodérmica, como hepatócitos e neurônios. Até o presente momento, os resultados dos estudos iniciais são animadores em relação ao uso de células-tronco mesenquimais e uso eficaz destas no reparo de tecidos retinianos lesados.

Stem cells have been studied in several fields of Medicine, and their applications are not too far from the clinical practice. Retinal impairment by neuronal death has been considered incurable due to the limited regenerative capacity of the central nervous system. The capacity of stem cells to regenerate tissues, as well as their plasticity makes them a potential source for retinal repair. The stem cells are a great promise for the therapy of inherited retinal disorders and retinal-neuronal degenerative diseases, such as retinitis pigmentosa and allied retinal dystrophies, which can result in blindness. Because of the accessibility, expansibility, and multipotentiality mesenchymal stem cells are expected to be useful for clinical applications, especially in regenerative medicine and tissue engineering. Mesenchymal stem cells are clonogenic, nonhematopoietic stem cells present in the bone marrow. Given the appropriate microenvironment, they could differentiate into cardiomyocytes or even into cells of nonmesodermal derivation including hepatocytes and neurons. So far, the results of a few studies are consistent with the belief that cell-based therapies using mesenchymal stem cells may be effective when it comes to retinal damaged tissue repair.