Mesenchymal stem cell therapy for equine tendinitis

Tendinitis is an important disease that leads to lameness and decreased performance in equine athletes and results in high costs associated with therapy due to a long recovery period and a high rate of recurrence. Although, several treatments for equine tendinitis have been described, few are effective in significantly improving the quality of the extracellular matrix and reducing the rate of recurrence. The use of cell therapy with mesenchymal stem cells (MSCs) derived from various sources has received much attention because of its therapeutic potential for equine tendinitis. In this paper, we review patents on stem cell therapy and the specific use of MSCs for the treatment of equine tendinitis. © 2013 Bentham Science Publishers.

Seedling morphology and development in the epiphytic cactus Epiphyllum phyllanthus (L.) Haw. (Cactaceae: Hylocereeae)

Seedling morphology is relevant in classification, taxonomy, and vegetation studies to understand plant life cycles, germination succession and requirements, and developmental progression. However, most morphological studies of seedlings lack analysis of organ anatomy, impeding the comprehension of series of development and establishment in a particular environment. Here, we have taken a traditional anatomical approach to examine the stages of seedling development in Epiphyllum phyllanthus, a holo-epiphytic cactus of tribe Hylocereeae. The goals were 1) to offer a comprehensive description of growth series in E. phyllanthus seedlings based on morphological and anatomical analysis and 2) to examine the initial growth phases in the life cycle of this species to identify organ development and understand their adaptive significance in relation to seedling establishment. Our results include descriptions of seed morphology, embryonic features, and seedling vascularization pattern in the root, hypocotyl, cotyledons, and epicotyl. The morphological and developmental patterns in E. phyllanthus seedlings have potential phylogenetic and ontogenetic implications in the Cactaceae. Characters such as the presence of mucilage on the seed coat, the lack of seed operculum, and large cotyledons in E. phyllanthus are comparable to basal cacti, but the root anatomy is more similar to columnar relatives. At the familial level, there is an apparent trend in decreasing number of phloem and xylem poles in the stele of primary root, correlated with degree of specialization and advanced phylogenetic position: tetrarch to septarch-octarch in basal lineages, tetrarch Cereus-type in columnar species, to the diarch vascular system in Rhipsalideae and some species with cylindric/globose stem. © Torrey Botanical Club.

Platelet lysate 3D scaffold supports mesenchymal stem cell chondrogenesis: An improved approach in cartilage tissue engineering

Articular lesions are still a major challenge in orthopedics because of cartilage's poor healing properties. A major improvement in therapeutics was the development of autologous chondrocytes implantation (ACI), a biotechnology-derived technique that delivers healthy autologous chondrocytes after in vitro expansion. To obtain cartilage-like tissue, 3D scaffolds are essential to maintain chondrocyte differentiated status. Currently, bioactive 3D scaffolds are promising as they can deliver growth factors, cytokines, and hormones to the cells, giving them a boost to attach, proliferate, induce protein synthesis, and differentiate. Using mesenchymal stem cells (MSCs) differentiated into chondrocytes, one can avoid cartilage harvesting. Thus, we investigated the potential use of a platelet-lysate-based 3D bioactive scaffold to support chondrogenic differentiation and maintenance of MSCs. The MSCs from adult rabbit bone marrow (n=5) were cultivated and characterized using three antibodies by flow cytometry. MSCs (1×105) were than encapsulated inside 60μl of a rabbit platelet-lysate clot scaffold and maintained in Dulbecco's Modified Eagle Medium Nutrient Mixture F-12 supplemented with chondrogenic inductors. After 21 days, the MSCs-seeded scaffolds were processed for histological analysis and stained with toluidine blue. This scaffold was able to maintain round-shaped cells, typical chondrocyte metachromatic extracellular matrix deposition, and isogenous group formation. Cells accumulated inside lacunae and cytoplasm lipid droplets were other observed typical chondrocyte features. In conclusion, the usage of a platelet-lysate bioactive scaffold, associated with a suitable chondrogenic culture medium, supports MSCs chondrogenesis. As such, it offers an alternative tool for cartilage engineering research and ACI. © 2013 Informa UK Ltd.

Immunophenotypic, immunocytochemistry, ultrastructural, and cytogenetic characterization of mesenchymal stem cells from equine bone marrow

The aim of this study was to isolate, culture, and characterize mesenchymal stem cells (MSCs) from horse bone marrow (BM) using the techniques of flow cytometry, immunocytochemistry, cytogenetics, and electron microscopy. Immunophenotypic analysis revealed the presence of MSCs with high expression of the CD90 marker, lower expression of the CD44 marker, and absent expression of the CD34 marker. In assays of differentiation, the positive response to osteogenic (OST), chondrogenic (CDG), and adipogenic (ADP) differentiation signals was observed and characterized by deposition of calcium-rich extracellular matrix (OST), proteoglycans and collagen II (CDG) and intracellular deposition of fat drops (ADP). In immunocytochemical characterization, MSCs were immunopositive for CD44, vimentin, and PCNA, and they were negative for CD13. In the ultrastructural analysis of MSCs, the most outstanding characteristic was the presence of rough endoplasmic reticulum with very dilated cisterns filled with a low electrodensity material. Additionally, MSCs had normal karyotypes (2n=64) as evidenced by cytogenetic analysis, and aneuploidy in metaphase was not observed. The protocols for isolating, culturing, and characterizing equine MSCs used in this study were shown to be appropriate for the production of a cell population with a good potential for differentiation and without aneuploidy that can be used to study future cellular therapies. © 2013 Wiley Periodicals, Inc.

Anatomy of scapes, bracts, and leaves of Paepalanthus sect. Diphyomene (Eriocaulaceae, Poales) and its taxonomic implications

Paepalanthus sect. Diphyomene comprises 18 species with a convoluted taxonomic history. Aiming to correlate anatomical structures with the systematics of this group and its relatives, we studied the anatomy of scapes, reproductive axis bracts, and leaves of 20 Paepalanthus species. Bracts and leaves show differences in epidermal cell thickening; mesophyll width; vascular bundle arrangement; presence or absence of a hypodermis; types of cells in the vascular bundle sheath extensions; margin shape and composition; and presence or absence of aquiferous parenchyma. Scapes differ in contour, rib number, and pith size. Some diagnostic characters found are presence of aquiferous parenchyma and absence of vascular bundle sheath extensions in leaves of P. urbanianus; vascular bundles decreasing in size towards the margin of leaves and bracts, and scapes with a triangular contour in P. flaccidus; scapes with nine ribs in P. acanthophyllus and ten in P. macer. All anatomical features are summarized in tables. These results aid in the identification and characterization of the species of P. sect. Diphyomene. They also support the current section circumscription, reinforcing the relevance of the anatomical characters in order to define natural groups. © 2012 The New York Botanical Garden.

Tissue-engineered blood vessel substitute by reconstruction of endothelium using mesenchymal stem cells induced by platelet growth factors

Background: Cardiovascular diseases remain leaders as the major causes of mortality in Western society. Restoration of the circulation through construction of bypass surgical treatment is regarded as the gold standard treatment of peripheral vascular diseases, and grafts are necessary for this purpose. The great saphenous vein is often not available and synthetic grafts have their limitations. Therefore, new techniques to produce alternative grafts have been developed and, in this sense, tissue engineering is a promising alternative to provide biocompatible grafts. This study objective was to reconstruct the endothelium layer of decellularized vein scaffolds, using mesenchymal stem cells (MSCs) and growth factors obtained from platelets. Methods: Fifteen nonpregnant female adult rabbits were used for all experiments. Adipose tissue and vena cava were obtained and subjected to MSCs isolation and tissue decellularization, respectively. MSCs were subjected to differentiation using endothelial inductor growth factor (EIGF) obtained from human platelet lysates. Immunofluorescence, histological and immunohistochemical analyses were employed for the final characterization of the obtained blood vessel substitute. Results: The scaffolds were successfully decellularized with sodium dodecyl sulfate. MSCs actively adhered at the scaffolds, and through stimulation with EIGF were differentiated into functional endothelial cells, secreting significantly higher quantities of von Willebrand factor (0.85 μg/mL; P < .05) than cells cultivated under the same conditions, without EIGF (0.085 μg/mL). Cells with evident morphologic characteristics of endothelium were seen at the lumen of the scaffolds. These cells also stained positive for fascin protein, which is highly expressed by differentiated endothelial cells. Conclusions: Taken together, the use of decellularized bioscaffold and subcutaneous MSCs seems to be a potential approach to obtain bioengineered blood vessels, in the presence of EIGF supplementation. © 2013 Society for Vascular Surgery.

Equine tendonitis therapy using mesenchymal stem cells and platelet concentrates: A randomized controlled trial

Introduction. Tendon injury is a major cause of lameness and decreased performance in athletic equines. Various therapies for tendonitis have been described; however, none of these therapies results in complete tissue regeneration, and the injury recurrence rate is high even after long recovery periods involving rest and physiotherapy. Methods. A lesion was induced with collagenase gel in the superficial digital flexor tendon in the center portion of the metacarpal region of eight equines of mixed breed. After two weeks, the lesions of the animals in the treated and control groups were treated through the intralesional administration of mesenchymal stem cells derived from adipose tissue (adMSCs) suspended in platelet concentrate (PC) and with phosphate buffered saline (PBS), respectively. Serial ultrasound analyses were performed every two weeks. After 16 weeks of therapy, a biopsy was performed for histopathological, immunohistochemical and gene expression (type I collagen (COL1A1), type III collagen (COL3A1), tenascin-C (TNC), tenomodulin (TNMD), and scleraxis (SCX)) analyses. Results: Differences in the ultrasound and histopathological analyses were observed between the groups. Improved results were reported in the group treated with adMSCs suspended in PC. There was no difference in the gene expression levels observed after the different treatments. The main results observed from the histopathological evaluation of the treated group were as follows: a prevention of the progression of the lesion, a greater organization of collagen fibers, and a decreased inflammatory infiltrate. A lack of progression of the lesion area and its percentage was observed in the ultrasound image, and increased blood flow was measured by Power Doppler. Conclusions: The use of adMSCs combined with PC for the therapy of experimentally induced tendonitis prevented the progression of the tendon lesion, as observed in the ultrasound examination, and resulted in a greater organization and decreased inflammation, as observed in the histopathological evaluation. These data demonstrate the therapeutic potential of this therapy for the treatment of equine tendonitis. © 2013 Carvalho et al.; licensee BioMed Central Ltd.

Source-driven remobilizations of nutrients within stem wood in Eucalyptus grandis plantations

Nutrient remobilizations in tree ligneous components have been little studied in tropical forests. A complete randomized block design was installed in Brazilian eucalypt plantations to quantify the remobilizations of phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) within stem wood. Three treatments were studied: control with neither K nor Na addition (C), 3 kmol ha-1 K applied (+K), and 3 kmol ha-1 Na applied (+Na). Biomass and nutrient contents were measured in the stem wood of eight trees destructively sampled at 1, 2, 3 and 4 years after planting in each treatment and annual rings were localized on discs of wood sampled every 3 m in half of the trees. Chemical analyses and wood density measurements were performed individually for each ring per level and per tree sampled. Nutrient remobilizations in annual rings were calculated through mass balance between two successive ages. Our results show that nutrient remobilizations within stem wood were mainly source-driven. Potassium and Na additions largely increased their concentration in the outer rings as well as the amounts remobilized in the first 2 years after the wood formation. The amount of Na remobilized in annual rings was 15 % higher in +Na than in +K the fourth year after planting despite a 34 % higher production of stem wood in +K leading to a much higher nutrient sink. A partial substitution of K by Na in the remobilizations within stem wood might contribute to enhancing Eucalyptus grandis growth in K-depleted soils. © 2013 Springer-Verlag Berlin Heidelberg.

Interaction between mesenchymal stem cells and Ti-30Ta alloy after surface treatment

In this study, in vitro cytocompatibility was investigated in the Ti-30Ta alloy after two kinds of surfaces treatments: alkaline and biomimetic treatment. Each condition was evaluated by scanning electron microscopy/energy-dispersive X-ray spectroscopy. Cellular adhesion, viability, protein expression, morphology, and differentiation were evaluated with Bone marrow stromal cells (MSCs) to investigate the short and long-term cellular response by fluorescence microscope imaging and colorimetric assays techniques. Two treatments exhibited similar results with respect to total protein content and enzyme activity as compared with alloy without treatment. However, it was observed improved of the biomineralization, bone matrix formation, enzyme activity, and MSCs functionality after biomimetic treatment. These results indicate that the biomimetic surface treatment has a high potential for enhanced osseointegration. © 2013 Wiley Periodicals, Inc.

Produção de plantas de arroz sob a ação de silício e nitrogênio

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

Características anatômicas foliares e controle químico em pós-emergência de Brachiaria decumbens e Brachiaria plantaginea

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

Anatomia comparada do lenho de raiz e caule de Lippia salviaefolia Cham. E Aegiphila sellowiana Cham.(Verbenaceae) de área de cerrado do Estado de São Paulo

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Stem cell factor estimula células da musculatura lisa da traqueia a produzir TGF-β, FGF-2 E CCL3/MIP-1α

Pós-graduação em Ciências Fisiológicas - FOA

Desenvolvimento de larenjeira 'Pera' Citrus Sinensis (L.) Osbeck enxertada em limoeiro 'Cravo' (Citrus Limonia) e cultivada em pó de balsalto

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Proteína capsidial do Rupestris stem pitting-associated vírus: seqüenciamento do gene, expressão em Escherichia coli, purificação e produção de anti-soro policlonal

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