Naisandra Bezerra da Silva

BC nanofibres: In vitro study of genotoxicity and cell proliferation

Nanomaterials have unusual properties not found in the bulk materials, which can be exploited in numerous applications such as biosensing, electronics, scaffolds for tissue engineering, diagnostics and drug delivery. However, research in the past few years has turned up a range of potential health hazards, which has given birth to the new discipline of nanotoxicology. Bacterial cellulose (BC) is a promising material for biomedical applications, namely due its biocompatibility. Although BC has been shown not to be cytotoxic or genotoxic, the properties of isolated BC nanofibres (NFs) on cells and tissues has never been analysed. Considering the toxicity associated to other fibre-shaped nanoparticles, it seems crucial to evaluate the toxicity associated to the BC-NFs. In this work, nanofibres were produced from bacterial cellulose by a combination of acid and ultrasonic treatment. The genotoxicity of nanofibres from bacterial cellulose was analysed in vitro, using techniques previously demonstrated to detect the genotoxicity of fibrous nanoparticles. The results from single cell gel electrophoresis (also known as comet assay) and the Salmonella reversion assays showed that NFs are not genotoxicity under the conditions tested. A proliferation assay using fibroblasts and CHO cells reveals a slight reduction in the proliferation rate, although no modification in the cell morphology is observed.

Evaluating the possible genotoxic, mutagenic and tumor cell proliferation-inhibition effects of a non-anticoagulant, but antithrombotic algal heterofucan

Fucan is a term used to denominate a family of sulfated polysaccharides rich in L‐fucose. They are extracted mainly from brown seaweeds and echinoderms. The brown seaweed Spatoglossum schröederi (Dictyotaceae) synthesizes three heterofucans named A, B and C. Our research group purified a non‐anticoagulant heterofucan (fucan A) which displays antithrombotic activity in vivo. However, its in vitro toxicity has yet to be determined. This work presents the evaluation of the potential cytotoxicity, mutagenicity and genotoxicity of this fucan. After 48 h incubation fucan A cytotoxicity was determinate using MTT assay. Tumor‐cell (HeLa, PC3, PANC, HL60) proliferation was inhibited 2.0–43.7%; at 0.05–1 mg ml−1 of the heterofucan, the 3T3 non‐tumor cell line proliferation was also inhibited (3.3–22.0%). On the other hand, the CHO tumorigenic and RAW non‐tumor cell lines proliferation were not affected by this molecule (0.05–1 mg ml−1). We observed no mutagenic activity in Salmonella reversion assay when bacterial strains TA97a, TA98, TA100 and TA102 (with and without S9) were used. Comet assay showed that fucan A had no genotoxic effect (from 20 to 1000 µg ml−1) on CHO cells. In conclusion, this study indicates that the S. schröederi fucan A was not found to be genotoxic or mutagenic compound; thus it could be used in new antithrombotic drug development. Copyright

Effect of titanium surface modified by plasma energy source on genotoxic response in vitro

Titanium (Ti) is currently the most widely used material for the manufacture of orthopedic and dental implants. Changes in the surface of commercial pure Ti (cp Ti) can determine the functional response of cells, and is therefore a critical factor for the success of the implant. However, the genotoxicity of titanium surfaces has been poorly studied. Thus, the purpose of this study was to evaluate the genotoxic potential of a new titanium surface developed by plasma treatment using argon-ion bombardment and compare it with an untreated titanium surface. Accordingly, comet assay, analysis of chromosomal aberrations (CAs), and Cytokinesis Block Micronucleus (CBMN) assay were carried out, using CHO-K1 (Chinese hamster ovary) cells grown on both titanium surfaces. Our results show that the untreated titanium surface caused a significant increase in % tail moment, in the number of cells with CAs, tetraploidy, micronucleus frequency, and other nuclear alterations when compared with the negative control and with the plasma-treated titanium surface. This difference may be attributed to increased surface roughness and changes in titanium oxide layer thickness.

Evaluation of acute and subchronic toxicity of a non-anticoagulant, but antithrombotic algal heterofucan from the Spatoglossum schröederi in Wistar rats

Fucan is a term used to denominate a family of sulfated polysaccharides rich in L-fucose. The brown alga Spatoglossum schroederi, Dictyotaceae, synthesizes three heterofucans named A, B, and C. Fucan A is a non-anticoagulant heterofucan which possesses potent antithrombotic (in vivo) and antiproliferative (in vitro) activities. However, its toxicity in vivo has not been determined. The present study examined the acute and subchronic toxicity of the fucan A in Wistar rats after subcutaneous administration. After that, the animals were killed and examined. The results showed in the acute study that fucan A did not cause general adverse effects and mortality in the concentrations 0, 20, 100, 1000, and 2000 µg/g body weight per rat for seven days. Regarding the subchronic study, the data showed that the fucan A did not cause any change in hematological and biochemistry parameters, as well as in the morphology, and in the size of the rats organs analyzed at a concentration of 20 µg/g body weight per rat during a 62-day period. In conclusion, this study indicates this heterofucan is a compound with potential pharmacological value that has no toxicity in vivo.

Morfologia do tubo digestório da tartaruga verde (Chelonia mydas)

The morphology of the digestive tube of the green turtle (Chelonia mydas) was evaluated. Nine young e 1 adult turtles were analyzed. The digestive organs withdrew occurred after plastron opening. The lengths description and measurement from each organ [esophagus, stomach, small intestine (SI) and large intestine (LI)] were made with digestive tube opened. The results showed that in both the adult and young animals, the esophagus from all species were marked by the presence of pointed papillae on internal mucous. The stomach presented saclike shape with blind fund. In the SI, reticular pleats in the duodenal mucous membrane have been observed, while the jejune and the ileum showed rectilinear longitudinal pleats. The LI was marked by the alternation of arched areas (haustra or sacculations) and narrowings. The microscopy of digestive tube from C. mydas revealed esophagus with folded mucous covered by keratinized and stratified squamous epitheliums. The stomach was divided in regions: cardiac, fundic e pyloric which differed as to the number of glands and the muscle layers arrangement. The SI presented marked by microvillus, and LI by folded mucous with glands the lamina propria. The morphology of the tube digestive of the green turtle showed to be adapted to your eating habits. A long the digestive tract and specializations provide greater area of absorption and more efficiency in digestion in this species.

Anatomy of the digestive tube of sea turtles (Reptilia: Testudines)

This study characterized the morphology of the digestive tube of five species of sea turtles. We used specimens found dead along the coast of the state Rio Grande do Norte, as well as specimens accidentally killed as a result of pelagic longline fishing. Nineteen animals of the following species were analyzed: Chelonia mydas (Linnaeus, 1758) (n = 9), Lepidochelys olivacea (Eschscholtz, 1829) (n = 6), Caretta caretta (Linnaeus, 1758) (n = 2), Eretmochelys imbricata (Linnaeus, 1766) (n = 1) and Dermochelys coriacea (Vandelli, 1761) (n = 1). After opening the plastron, we removed the digestive organs and described the external and internal morphology of each organ. The esophagus of all species had pointed papillae on the mucosa. The stomach varied in shape among species. Differences were found in the mucosa of the small intestine. It was reticular in the duodenum, and longitudinal rectilinear in the jejunum/ileum. In all species an alternation of saccular and narrow regions was observed in the large intestine. The exception was D. coriacea, in which the mucosa of the entire large intestine had irregularly distributed folds. The pattern of the esophagus was the same in all species. The morphology of the stomach differed among species, and these differences reflect their diets. In addition, the distribution pattern of the folds on the mucosa of the small intestine varied between regions of the intestine and among species.

Biological activities of the sulfated polysaccharide from the vascular plant Halodule wrightii

A sulfated polysaccharide (SPSG) was successfully isolated from seagrass Halodule wrightii Asch., Cymodoceaceae, and its antioxidant and anticoagulant activities were investigated. The data presented here showed that the SPSG is a 11 kDa sulfated heterogalactan with a sulfatation degree of 20.63% and it also contains glucose and xylose. SPSG antioxidant activities were evaluated using several in vitro assays and the anticoagulant activity was evaluated by aPTT and PT tests. These assays suggested that the SPSG possessed remarkable antioxidant properties in different in vitro assays and an outstanding anticoagulant activity 2.5-fold higher than that of heparin Clexane® in the aPTT test. This data represents the first reported on the sulfated polysaccharide biological activities from seagrass. These results indicate that SPSG can be considered in the future as a drug utilized in treating diseases from these systems.

Morphological features and vascularization study of caprine cyclic corpus luteum

Miranda-Moura M.T.M., Fonseca V.U., Silva N.B., Freitas M.L., Almeida O.B., Rocha H.A.O., Papa P.C. & Moura C.E.B. 2010. Morphological features and vascularization study of caprine cyclic corpus luteum. Pesquisa Veterinaria Brasileira 30(4):351-357. Departamento de Morfologia, Centro de Biociencias, Universidade Federal do Rio Grande do Norte, Av. Salgado Filho s/n, Campus Lagoa Nova, Natal, RN 59072- 970, Brazil. E-mail: mouraeduard@cb.ufrn.br Corpus luteum is a temporary endocrine gland that regulates either the estrous cycle and pregnancy. It presents extreme dependency on the adequate blood supply. This work aims to evaluate goat corpus luteum (CL) vascular density (VD) over the estrous cycle. For that purpose, 20 females were submitted to estrus synchronization/ovulation treatment using a medroxyprogesterone intra-vaginal sponge as well as intramuscular (IM) application of cloprostenol and equine chorionic gonadotrophine (eCG). After sponge removal, estrus was identified at about 72hs. Once treatment was over, female goats were then subdivided into 4 groups (n=5 each) and slaughtered on days 2, 12, 16 and 22 after ovulation (p.o). Ovaries were collected, withdrawn and weighted. CL and ovaries had size and area recorded. Blood samples were collected and the plasma progesterone (P4) was measured through RIA commercial kits. The VD was 24.42±6.66, 36.26±5.61, 8.59±2.2 and 3.97±1.12 vessels/mm 2 for days 2, 12, 16 and 22 p.o, respectively. Progesterone plasma concentrations were 0.49±0.08, 2.63±0.66, 0.61±0.14 and 0.22±0.04ng/ml for days 2, 12, 16 e 22 p.o, respectively. Studied parameters were affected by the estrous cycle phase. Values greater than 12 p.o were observed. In the present work we observed that ovulation occurred predominantly in the right ovary (70% of the animals), which in turn presented bigger measures than the contra lateral one. There is a meaningful relationship between the weight and size of the ovary and these of CL (r=0.87, r=0.70, respectively, p<0.05). It is possible to conclude that morphology of goats ovaries and plasma progesterone concentration changed according to estrous cycle stages. We propose these parameters can be used as indicators of CL functional activity.

MC3T3‐E1 Cells Behavior on Surfaces Bombarded by Argon Ions in Planar Cathode Discharge

To evaluate the effect of topography in nanoscale, titanium surfaces were bombarded by argon ions (a chemically inert gas), in an atmosphere of plasma. The effects of surface parameters on morphology, adhesion, proliferation, and MC3T3-E1 preosteoblasts differentiation were analyzed. Nontreated (smooth) surfaces were used as a control. The levels of average roughness (Ra) observed in bombarded and smooth titanium surfaces were of 95 and 14 nm, respectively. The wettability increased on treated surfaces. The number of attached cells (30 and 60 min) was significantly higher on the bombarded surface. The cell proliferation after 3 and 7 days was also significantly higher on the ion-bombarded surface. In addition, the ALP activity and expression of osteocalcin were higher in cells grown on the treated surface. The results showed that bombardment with argon ions increased the roughness and the wettability of the Ti surface, promoting a significant increase in the adhesion, proliferation, and differentiation of preosteoblasts.

In Vivo Evaluation of the Antioxidant Activity and Protective Action of the Seaweed Gracilaria birdiae

The red seaweed Gracilaria birdiae (GB) is farmed and used as food in northeast Brazil. However, the economic potential of this seaweed has been explored little. To enable direct consumption and/or product diversification from GB, it is necessary to evaluate its effect in vivo. In this study, the food of mice was improved with the addition of GB. After 21 days, the consumption of seaweed reduced the weight gain and blood glucose levels in mice. In addition, it increased the trolox equivalent antioxidant capacity and glutathione reductase and catalase levels compared to those of the control group. In addition, some mice also received carbon tetrachloride (CCl4). In this case, histological, enzymatic, and antioxidant tests showed that the seaweed could protect animals from damage caused by this toxic agent. In addition, GB aqueous extract (AE) inhibited 50% of 3T3-L1 cell differentiation into adipocytes, whereas GB ethanolic extract was not effective. AE is composed mainly of sulfated polysaccharides. The results of the present study indicate that the alga GB protected the mice from CCl4-induced damage, indicating that the seaweed exhibits protective action in vivo. In addition, GB decreased the animal weight gain, which was mainly due to the action of the sulfated polysaccharides synthesized by this seaweed.