Equine mesenchymal stem cells from bone marrow, adipose tissue and umbilical cord: immunophenotypic characterization and differentiation potential

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Bone Marrow'S Harvest in The Coxal Tuberosity for Isolation and Culture of Mesenchymal Stem Cells of Buffaloes (Bubalus Bubalis)

Several studies with mesenchymal stem cells (MSCs) have been developed in many species because of its ability to differentiate into other mesoderm lineages, capacity of self-regeneration, low immunogenicity, paracrine, anti-inflamatory, immunomodulatory and antiapoptotic effects which make then a promissory source to be used in therapeutic strategies. The aim of this study is to report the technique of harvest of bone marrow (BM) in the coxal tuberosity (CT) of buffaloes. For this, the animals were selected, identified and contained in a stock. Then trichotomy was performed in the region corresponding to the CT. After identifying the anatomic site it was performed antisepsis, local anesthetic block and introduction of a myelogram's needle (Lang(R)) for BM aspiration. Once the needle was firmly fixed in the CT, the mandril was removed and proceeded to BM aspiration with a syringe (20 mL) containing 1 ml of heparin at 1000 IU / mL and 1 mL of PBS. After the collection, each sample collected was manually homogenized, identified and referred to the LRACT - FMVZ / UNESP-BRAZIL for the correct processing. The anatomical site tested showed to be an alternative site of harvest of BM once provided the appropriate isolation and culture of the mononuclear fraction. Moreover, the procedure was performed without difficulty and with great security. Based on this, it can be conclude that CT is an excellent anatomical site for isolation and culture of MSCs and the proposed technique is viable and feasible to be held in buffaloes.

Isolation, Culture and Differentiation of Buffaloes Bone Marrow Mesenchymal Stem Cells Obtained from the Coxal Tuberosity

Currently, much attention has been devoted to the renewal of knowledge about Stem Cells and Cell Therapy in domestic species. In this sense, the present work aimed to develop a methodology for collecting, processing and cultivation of mesenchymal stem cells obtained from bone marrow of coxal tuberosity in buffaloes. The collection was performed using a Komiyashiki needle, which was introduced in the coxal tuberosity and the bone marrow aspirated into a heparinized syringe with the aid of negative pressure. Directly after collection samples were processed at the laboratory at FMVZ - UNESP. The samples took approximately 32 days to reach 80% confluence, when the first passage and differentiation was performed. To confirm the mesenchymal origin, cells were induced to differentiate into adipogenic and osteogenic lineages. Samples showed morphological changes during differentiation protocol, but not all presented production of extracellular deposits of calcium or intracellular fat droplets, observed after staining with Alizarin Red and Oil Red respectively. Compared with the material obtained from other species and processed in the same laboratory, the primary culture was longer. Therefore, more studies are needed to standardize the age of animals used and to test other inducers of cell differentiation.

Root system and root and stem base organic reserves of pasture Tanzania grass fertilizer with nitrogen under grazing

The goal of this study was to evaluate the concentrations of non-structural carbohydrate (NSC) and of total nitrogen (N), as well as, to evaluate the root system in Tanzania-grass pastures fertilized with doses of urea in fall, spring and summer. The experiment was conducted at the Experimental Farm of Iguatemi, Maringa, Parana, Brazil, from March 2007 to March 2008. The experimental design was complete random blocks with subplots and four repetitions. The plots showed doses of N (50, 100 e 150 kg ha(-1) of N) plus the control (no N fertilization), and the subplots the season of the year. Root samples were taken at depths of 0-10, 10-20 and 20-40 cm. Root biomass showed a trend for mass accumulation up to a dosage of 100 kg ha(-1) for all seasons evaluated. Also, about 80% of the root system of Tanzaniagrass plants was found on the 0-10 cm layer for all dosages of N. Nitrogen fertilizer above 100 kg ha(-1) may foster fast forage plant growth reducing its NSC root storage capacity although favoring NSC and total N storage at stem base. NSC and total N concentrations were highest in fall, demonstrating that its usage is greater in spring due to the weather conditions being favorable to plant growth. In the regrowth, the largest reserve of total N was at the 0-10 cm root layer and the largest NSC reserve is at stem base.

Bone marrow mesenchymal stem cells stimulated by bFGF up-regulated protein expression in comparison with periodontal fibroblasts in vitro

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

Influence of Tapered and External Hexagon Connections on Bone Stresses Around Tilted Dental Implants: Three-Dimensional Finite Element Method With Statistical Analysis

Background: The purpose of this study is to analyze the tension distribution on bone tissue around implants with different angulations (0 degrees, 17 degrees, and 30 degrees) and connections (external hexagon and tapered) through the use of three-dimensional finite element and statistical analyses.Methods: Twelve different configurations of three-dimensional finite element models, including three inclinations of the implants (0 degrees, 17 degrees, and 30 degrees), two connections (an external hexagon and a tapered), and two load applications (axial and oblique), were simulated. The maximum principal stress values for cortical bone were measured at the mesial, distal, buccal, and lingual regions around the implant for each analyzed situation, totaling 48 groups. Loads of 200 and 100 N were applied at the occlusal surface in the axial and oblique directions, respectively. Maximum principal stress values were measured at the bone crest and statistically analyzed using analysis of variance. Stress patterns in the bone tissue around the implant were analyzed qualitatively.Results: The results demonstrated that under the oblique loading process, the external hexagon connection showed significantly higher stress concentrations in the bone tissue (P < 0.05) compared with the tapered connection. Moreover, the buccal and mesial regions of the cortical bone concentrated significantly higher stress (P < 0.005) to the external hexagon implant type. Under the oblique loading direction, the increased external hexagon implant angulation induced a significantly higher stress concentration (P = 0.045).Conclusions: The study results show that: 1) the oblique load was more damaging to bone tissue, mainly when associated with external hexagon implants; and 2) there was a higher stress concentration on the buccal region in comparison to all other regions under oblique load.

The root systems of permian arborescent sphenopsids: evidence from the northern and southern hemispheres

Anatomically preserved calamitalean trunks are described from the Permian fossil forests of Chemnitz, Germany, and Tocantins, central-north Brazil. Several trunk bases were found in situ, still rooting in their former substrate or in parautochthonous sediments and revealing multiple organic connections between stems and roots. The new evidence of several free-stemmed Permian calamitaleans from different fossil lagerstatten and different taphonomic modes from the Northern and Southern hemispheres has implications for understanding calamite growth and challenges the universal validity of the reconstruction of rhizome-bearing woody trees. Whereas the stems belong to different species of the widely distributed genus Arthropitys GOEPPERT 1864, among them the generitype A. bistriata (COTTA) emend. RoSSLER, FENG & NOLL 2012 the attached roots represent the largest calamite roots ever found and incorporate a broad spectrum of preservational forms and ontogenetic stages. The latter are represented by the root genera Astromyelon WILLIAMSON 1878, Myriophylloides HICK & CASH 1881 and Asthenomyelon LEISTIKOW 1962 that were evidenced for the first time from Chemnitz, the type locality of Arthropitys and Calamitea (COTTA) emend. ROSSLER & NOLL 2007. Branched, stem-borne, adventitious root systems exhibit similar architectures, arise from different nodes of the lowermost trunks and anchor the trees in' different substrates. Developmental features were analysed in first- to third-order roots, which possess clearly-defined concentric tissue zones: epidermis/periderm, cortex, endodermis and central vascular tissue with or without pith. First-order roots, in particular, show considerable secondary growth. Numerous zones of concentric density variation in the secondary xylem indicate some kind of seasonality in the early Permian environments.

Novel Chemically Modified Bacterial Cellulose Nanocomposite as Potential Biomaterial for Stem Cell Therapy Applications

Bacterial cellulose (BC) has become established as a remarkably versatile biomaterial and can be used in a wide variety of applied scientific applications, especially for medical devices. In this work, the bacterial cellulose fermentation process is modified by the addition of hyaluronic acid and gelatin (1% w/w) to the culture medium before the bacteria is inoculated. Hyaluronic acid and gelatin influence in bacterial cellulose was analyzed using Transmission Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). Adhesion and viability studies with human dental pulp stem cells using natural bacterial cellulose/hyaluronic acid as scaffolds for regenerative medicine are presented for the first time in this work. MTT viability assays show higher cell adhesion in bacterial cellulose/gelatin and bacterial cellulose/ hyaluronic acid scaffolds over time with differences due to fiber agglomeration in bacterial cellulose/gelatin. Confocal microscopy images showed that the cell were adhered and well distributed within the fibers in both types of scaffolds.

ANTIDIARRHEAL MECHANISM AND IONIC PROFILE OF CARPOLOBIA LUTEA ETHANOLIC STEM-BARK EXTRACT IN RATS

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

Evaluation of the acidity in the juice from different parts of the stem of sugarcane, cycle 2010/2011

The acidity levels present in the juice of sugarcane can cause problems in its processing. This study aimed to compare the values of total, volatile and fixed acidity in the juice of the stem nodes and internodes of sugarcane. The experimental design was a completely randomized, 3x2 factorial arrangement with four replications, using three varieties of sugarcane (IAC91-1099; ISC95 and CTC7-5000) divided into node and internode. The stems were collected and separated in node and internode. After extraction of juice from each part, total, volatile and fix acidities were determined. The results were subjected to variance analysis the by F test and compared by the Tukey test at 5% probability. The cultivar CTC7 had the lowest levels of total and fixed acidity. Cultivar IAC91-1099 showed high levels of fixed acidity on both parts of the stem, and high volatile acidity at the node. In the stem, the values of volatile acidity showed no definite pattern between cultivars. The node has, in general, a higher fixed acidity, but some cultivars have shown opposite results.

Comparative wood anatomy of root and stem of Citharexylum myrianthum (Verbenaceae)

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

Phototherapy up-regulates dentin matrix proteins expression and synthesis by stem cells from human-exfoliated deciduous teeth

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