Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8

In vitro Related Publications

8 Effect of Copper Ions Doped-Hydroxyapatite 3D Fiber Scaffold

Authors: Adil Elrayah, Jie Weng, Esra Suliman

Abstract:

The mineral in human bone is not pure stoichiometric calcium phosphate (Ca/P) as it is partially substituted by in organic elements. In this study, the copper ions (Cu2+) substituted hydroxyapatite (CuHA) powder has been synthesized by the co-precipitation method. The CuHA powder has been used to fabricate CuHA fiber scaffolds by sol-gel process and the following sinter process. The resulted CuHA fibers have slightly different microstructure (i.e. porosity) compared to HA fiber scaffold, which is denser. The mechanical properties test was used to evaluate CuHA, and the results showed decreases in both compression strength and hardness tests. Moreover, the in vitro used endothelial cells to evaluate the angiogenesis of CuHA. The result illustrated that the viability of endothelial cell on CuHA fiber scaffold surfaces tends to antigenic behavior. The results obtained with CuHA scaffold give this material benefit in biological applications such as antimicrobial, antitumor, antigens, compacts, filling cavities of the tooth and for the deposition of metal implants anti-tumor, anti-cancer, bone filler, and scaffold.

Keywords: In vitro, Hardness, Mechanical Properties, hydroxyapatite, copper ions, fiber scaffold

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7 Effect of Different Media and Mannitol Concentrations on Growth and Development of Vandopsis lissochiloides (Gaudich.) Pfitz. under Slow Growth Conditions

Authors: P. Inthima, A. Kongbangkerd, J. Linjikao

Abstract:

In vitro conservation of orchid germplasm provides an effective technique for ex situ conservation of orchid diversity. In this study, an efficient protocol for in vitro conservation of Vandopsis lissochiloides (Gaudich.) Pfitz. plantlet under slow growth conditions was investigated. Plantlets were cultured on different strength of Vacin and Went medium (½VW and ¼VW) supplemented with different concentrations of mannitol (0, 2, 4, 6 and 8%), sucrose (0 and 3%) and 50 g/L potato extract, 150 mL/L coconut water. The cultures were incubated at 25±2 °C and maintained under 20 µmol/m2s light intensity for 24 weeks without subculture. At the end of preservation period, the plantlets were subcultured to fresh medium for growth recovery. The results found that the highest leaf number per plantlet could be observed on ¼VW medium without adding sucrose and mannitol while the highest root number per plantlet was found on ½VW added with 3% sucrose without adding mannitol after 24 weeks of in vitro storage. The results showed that the maximum number of leaves (5.8 leaves) and roots (5.0 roots) of preserved plantlets were produced on ¼VW medium without adding sucrose and mannitol. Therefore, ¼VW medium without adding sucrose and mannitol was the best minimum growth conditions for medium-term storage of V. lissochiloides plantlets.

Keywords: Germplasm, Preservation, In vitro, vandopsis

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6 Morphological Interaction of Porcine Oocyte and Cumulus Cells Study on in vitro Oocyte Maturation Using Electron Microscopy

Authors: M. Areekijseree, M. Pumipaiboon, T. Chuen-Im, W. Pongsawat, C. Thepsithar, S. Sengsai

Abstract:

Morphological interaction of porcine cumulus-oocyte complexes (pCOCs) was investigated on in vitro condition using electron microscope (SEM and TEM). The totals of 1,923 oocytes were round in shape, surrounded by Zona pellucida with layer of cumulus cells ranging between 59.29-202.14 μm in size. They were classified into intact-, multi-, partial cumulus cell layer oocyte, and completely denuded oocyte, at the percentage composition of 22.80% 32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as intact- and multi cumulus cell layer oocytes were further culturing at 37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2 mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 μg/mL porcine follicle-stimulating hormone, 1 μg/mL LH, 1μg/mL estradiol with ethanol, and 50 μg/mL gentamycin sulfate. On electron microscope study, cumulus cells were found to stick their processes to secrete substance from the sac-shape end into Zona pellucida of the oocyte and also communicated with the neighboring cells through their microvilli on the beginning of incubation period. It is believed that the cumulus cells communicate with the oocyte by inserting the microvilli through this gap and embedded in the oocyte cytoplasm before secreting substance, through the sac-shape end of the microvilli, to inhibit primary oocyte development at the prophase I. Morphological changes of the complexes were observed after culturing for 24-44 h. One hundred percentages of the cumulus layers were expanded and cumulus cells were peeling off from the oocyte surface. In addition, the round-shape cumulus cells transformed themselves into either an elongate shape or a columnar shape, and no communication between cumulus neighboring cells. After 44 h of incubation time, diameter of oocytes surrounded by cumulus cells was larger than 0 h incubation. The effect of hormones in culture medium is exerted by their receptors present in porcine oocyte. It is likely that all morphological changes of the complexes after hormone treatment were to allow maturation of the oocyte. This study demonstrated that the association of hormones in M199 could promote porcine follicle activation in 44 h in vitro condition. This culture system should be useful for studying the regulation of early follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell technology.

Keywords: In vitro, cumulus cells, porcine oocyte, electron microscopy (SEM and TEM)

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5 Effect of Various Pollen Sources to Ability Fruit Set and Quality in ‘Long Red B’ Wax Apple

Authors: Nguyen Minh Tuan, Yen Chung-Ruey

Abstract:

By hand pollination was conducted to evaluated different pollen sources and their affects on fruit set and quality of wax apple. The following parameters were recorded: fruit set, seed set, fruit characteristics. Results showed that fruit set percentage with seed were significantly high in ‘Long Red B’ when ‘Black’, ‘Thyto’ were used as pollen parents. Pollen of ‘Black’, ‘Thyto’ resulted in high fruit weight, fruit diameter, fruit length, bigger flesh thickness, better total soluble solids as compared with other pollens. The observation of pollen-growth in vitro revealed that pollen germination at 15% sucrose concentration are required for optimum pollen germination with the high pollen germination were found in ‘Black’, ‘Thyto’. From the result, we concluded that ‘Black’, ‘Thyto’ were proved to be good pollinizers in ‘Long Red B’. Therefore, artificial cross-pollination using ‘Black’, ‘Thyto’ as pollinizers were strongly recommended for ‘Long Red B’ cultivar in wax apple orchard.

Keywords: In vitro, pollination, fruit quality, wax apple, pollen source

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4 Effects of a Methanol Fraction of the Leaves of Leonotis leonurus on the Blood Pressure and Heart Rate of Normotensive Male Wistar Rats

Authors: K. Obikeze, P. Mugabo, I. Green, D. Dietrich, A. Burger

Abstract:

Leonotisleonurus a shrub indigenous to Southern Africa is widely used in traditional medicine to treat a variety of conditions ranging from skin diseases and cough to epileptic fits and ‘heart problems’. Studies on the aqueous extract of the leaves have indicated cycloxegenase enzyme inhibitory activity and an antihypertensive effect. Five methanol leaf extract fractions (MLEa - MLEe) of L. leonurus were tested on anaesthetized normotensive male Wistar rats (AWR) and isolated perfused working rat hearts (IWH). Fraction MLEc (0.01mg/kg – 0.05mg/kg) induced significant increases in BP and HR in AWR and positive chronotropic and inotropic effects in IWH (1.0mg/ml – 5.0mg/ml). Pre-administration of atenolol (2.0mg/kg) and prazosin (60μg/kg) significantly inhibited MLEc effect on HR and MAP respectively in vivo, while atenolol (7.0mg/ml) pre-perfusion significantly inhibited MLEc effect in vitro. The hypertensive effect of MLEc is probably via β1agonism. Results also indicate the presence of multiple cardioactive compounds in L. leonurus.

Keywords: in vivo, In vitro, rat, Cardiovascular effect, isolated perfused working heart, Leonotis leonurus

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3 Extractable Heavy Metal Concentrations in Bottom Ash from Incineration of Wood-Based Residues in a BFB Boiler Using Artificial Sweat and Gastric Fluids

Authors: Risto Pöykiö, Hannu Nurmesniemi, Olli Dahl

Abstract:

The highest extractable concentration in the artificial sweat fluid was observed for Ba (120mg/kg; d.w.). The highest extractable concentration in the artificial gastric fluid was observed for Al (9030mg/kg; d.w.). Furthermore, the extractable concentrations of Ba (550mg/kg; d.w.) and Zn (400mg/kg: d.w.) in the bottom ash using artificial gastric fluid were elevated. The extractable concentrations of all heavy metals in the artificial gastric fluid were higher than those in the artificial sweat fluid. These results are reasonable in the light of the fact that the pH of the artificial gastric fluid was extremely acidic both before (pH 1.54) and after (pH 1.94) extraction, whereas the pH of the artificial sweat fluid was slightly alkaline before (pH 6.50) and after extraction (pH 8.51).

Keywords: Waste, Heavy Metals, In vitro, ash, artificial fluid

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2 Cultivation of Thymus by In Vitro And Hydroponics Combined Method

Authors: E. Sargsyan, A. Vardanyan, L. Ghalachyan, S. Bulgadaryan

Abstract:

Our results showed that for the growth of qualitative seedling and vegetative raw material of ðó. marschallianus Willd. and T. serphyllum L. it is more profitable to use the in vitro and hydroponics combined method. In in vitro culture it is possible to do micro-propagation whole year with 98-99% rhizogenesis. 30000 micro-plants were obtained from one explant during 9 months. Hydroponic conditions provide the necessary microclimate for microplants where the survival rate without acclimatization was 93.3%. The essential oil content in hydroponic dry herb of both species in vegetative and blossom phase was 1.3% whereas in wild plants it was 1.2%, the content of extractive substances and vitamin C also exceeded wild plants. Our biochemical and radiochemical investigations indicated that the medicinal raw materials obtained from hydroponic and wild plants of Thymus species correspond to the demands of SPh XI, and the content of artificial radionuclides does not exceed the MACL.

Keywords: In vitro, Hydroponics, thymus, Micro-propagation

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1 Effect of Recombinant Human Follicle Stimulating Hormone on Meiotic Competence of In Vitro Grown Nili Ravi Buffalo Oocytes

Authors: Muhammad Ijaz Khan, Samina Jalali, Beenish Shahid, S. A. Shami, Muhammad Ikramullah

Abstract:

In the present study, the response of Nili Ravi buffalo oocytes to recombinant human follicle stimulating hormone (rhFSH) (Organon) on meiotic maturation in vitro was examined. Oocytes were matured in vitro in medium containing either 0 or 0.05 IU/ ml rhFSH and the stage of nuclear maturation recorded after 24 hours. The percentage of oocytes in the control group undergoing germinal vesicle breakdown (GVBD) observed after 24 hours of culture was 29 % whereas as in rhFSH group the percentage was 10 % were at this stage (P< 0.001).Thus in the presence of rhFSH, a significantly greater number of oocytes had progressed to the more advanced stages of nuclear maturation. Indeed, the maturation of GV (Germinal Vesicle) stage oocytes to the metaphase II (M II) stage after 24 hours was significantly (P< 0.0001) increased by the addition of rhFSH (82 % VS 47 %). The percentage of degenerated oocytes after 24 hours of culture was 24 % in control group, whereas in rhFSH group the percentage was 8 % after 24 hours. Degeneration of the oocytes after 24 hours was not significantly (P = 0. 9361) decreased.

Keywords: In vitro, buffalo, oocytes, recombinant FSH

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