Search results for: tissue engineering

Authors: Emma K. Sales, Nilda G. Butardo

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The project was undertaken to determine the effects of modified tissue culture protocols e.g. age of culture and hormone levels (2,4-D) in generating somaclonal variation. Moreover, the utility of molecular markers (SSR and MSAP) in sorting off types/somaclones were investigated.

Results show that somaclonal variation is in effect due to prolonged subculture and high 2,4-D concentration. The resultant variation was observed to be due to high level of methylation events specifically cytosine methylation either at the internal or external cytosine and was identified by methylation sensitive amplification polymorphism (MSAP).Simple sequence repeats (SSR) on the other hand, was able to associate a marker to a trait of interest.

These therefore, show that molecular markers can be an important tool in sorting out variation/mutants at an early stage.

Keywords: Methylation, MSAP, somaclones, SSR, subculture, 2, 4-D.

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Authors: Chung-Yao Yang, Lin-Ya Huang, Tang-Long Shen, J. Andrew Yeh

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The importance for manipulating an incorporated scaffold and directing cell behaviors is well appreciated for tissue engineering. Here, we developed newly nano-topographic oxidized silicon nanosponges capable of being various chemical modifications to provide much insight into the fundamental biology of how cells interact with their surrounding environment in vitro. A wet etching technique is exerted to allow us fabricated the silicon nanosponges in a high-throughput manner. Furthermore, various organo-silane chemicals enabled self-assembled on the surfaces by vapor deposition. We have found that Chinese hamster ovary (CHO) cells displayed certain distinguishable morphogenesis, adherent responses, and biochemical properties while cultured on these chemical modified nano-topographic structures in compared with the planar oxidized silicon counterparts, indicating that cell behaviors can be influenced by certain physical characteristic derived from nano-topography in addition to the hydrophobicity of contact surfaces crucial for cell adhesion and spreading. Of particular, there were predominant nano-actin punches and slender protrusions formed while cells were cultured on the nano-topographic structures. This study shed potential applications of these nano-topographic biomaterials for controlling cell development in tissue engineering or basic cell biology research.

Keywords: Nanosponge, Cell adhesion, Cell morphology

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Authors: C. Gallottini, W. Di Mari, A. Amaddeo, K. Barbaro, A. Dolci, G. Dolci, L. Gallottini, G. Barraco, S. Eramo

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A group of 10 dogs (group A) with Periodontal Disease in the third stage, were subjected to regenerative therapy of periodontal tissues, by use of nano hydroxy apatite (NHA). These animals induced by general anesthesia, where treated by ultrasonic scaling, root planning, and at the end by a mucogingival flap in which it was applied NHA. The flap was closed and sutured with simple steps. Another group of 10 dogs (group B), control group, was treated only by scaling and root planning. No patient was subjected to antibiotic therapy. After three months, a check was made by inspection of the oral cavity, radiography and bone biopsy at the alveolar level. Group A showed a total restitutio ad integrum of the periodontal structures, and in group B still mild gingivitis in 70% of cases and 30% of the state remains unchanged. Numerous experimental studies both in animals and humans have documented that the grafts of porous hydroxyapatite are rapidly invaded by fibrovascular tissue which is subsequently converted into mature lamellar bone tissue by activating osteoblast. Since we acted on the removal of necrotic cementum and rehabilitating the root tissue by polishing without intervention in the ligament but only on anatomical functional interface of cement-blasts, we can connect the positive evolution of the clinical-only component of the cement that could represent this perspective, the only reason that Periodontal Disease become a Cement Disease, while all other clinical elements as nothing more than a clinical pathological accompanying.

Keywords: Nanoidroxiaphatite, Parodontal Disease, Rigenerative Therapy.

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Authors: Rusudan Sujashvili, Ekaterine Bakuradze, Irina Modebadze, Davit Dekanoidze

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Extracellular ubiquitin in vivo effect on regenerative liver cells and liver histoarchitectonics has been studied. Experiments were performed on mature female white rats. Partial hepatectomy was made using the modified method of Higgins and Anderson. Standard histopathological assessment of liver tissue was used. Proliferative activity of hepatocytes was analyzed by colchicine mitotic index and immunohistochemical staining on ki67. We have found that regardless of number of injections and dose of extracellular ubiquitin liver histology has not been changed, so at tissue level no effect was observed. In vivo double injection of ubiquitin significantly decreases the mitotic activity at 32 hour point after partial hepatectomy. Thus, we can conclude that in vivo injected extracellular ubiquitin inhibits proliferative activity of hepatocytes in partially hepatectomyzed rats.

Keywords: Liver, regeneration, proliferation, ubiquitin.

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Authors: M. S. Nourbakhsh, M. E. Khosroshahi

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Laser soldering is based on applying some soldering material (albumin) onto the approximated edges of the cut and heating the solder (and the underlying tissues) by a laser beam. Endogenous and exogenous materials such as indocyanine green (ICG) are often added to solders to enhance light absorption. Gold nanoshells are new materials which have an optical response dictated by the plasmon resonance. The wavelength at which the resonance occurs depends on the core and shell sizes, allowing nanoshells to be tailored for particular applications. The purposes of this study was use combination of ICG and different concentration of gold nanoshells for skin tissue soldering and also to examine the effect of laser soldering parameters on the properties of repaired skin. Two mixtures of albumin solder and different combinations of ICG and gold nanoshells were prepared. A full thickness incision of 2×20 mm2 was made on the surface and after addition of mixtures it was irradiated by an 810nm diode laser at different power densities. The changes of tensile strength σt due to temperature rise, number of scan (Ns), and scan velocity (Vs) were investigated. The results showed at constant laser power density (I), σt of repaired incisions increases by increasing the concentration of gold nanoshells in solder, Ns and decreasing Vs. It is therefore important to consider the tradeoff between the scan velocity and the surface temperature for achieving an optimum operating condition. In our case this corresponds to σt =1800 gr/cm2 at I~ 47 Wcm-2, T ~ 85ºC, Ns =10 and Vs=0.3mms-1.

Keywords: Tissue soldering, gold nanoshells, indocyanine green, combination, tensile strength.

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Authors: Amr A. Fouad, Hamed A. Alwadaani, Iyad Jresat

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The present study investigated the protective effect of thymoquinone (TQ), against cadmium-induced kidney injury in rats. Cadmium chloride (1.2 mg Cd/kg/day, s.c.), was given for nine weeks. TQ treatment (40 mg/kg/day, p.o.) started on the same day of cadmium administration and continued for nine weeks. TQ significantly decreased serum creatinine, renal malondialdehyde and nitric oxide, and significantly increased renal reduced glutathione in rats received cadmium. Histopathological examination showed that TQ markedly minimized renal tissue damage induced by cadmium. Immunohistochemical analysis revealed that TQ markedly decreased the cadmium-induced expression of inducible nitric oxide synthase, tumor necrosis factor-α, cyclooxygenase-2, and caspase-3 in renal tissue. It was concluded that TQ significantly protected against cadmium nephrotoxicity in rats, through its antioxidant, antiinflammatory, and antiapoptotic actions.

Keywords: Thymoquinone, cadmium, kidney, rats.

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Authors: Amr A. Fouad, Waleed H. Albuali, Iyad Jresat

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The protective effect of thymoquinone (TQ) was investigated in rats exposed to testicular injury induced by sodium arsenite (10mg/kg/day, orally, for two days). TQ treatment (10mg/kg/day, intraperitoneal injection) was applied for five days, starting three day before arsenic administration. TQ significantly attenuated the arsenic-induced decreases of serum testosterone, and testicular reduced glutathione level, and significantly decreased the elevations of testicular malondialdehyde and nitric oxide levels resulted from arsenic administration. Also, TQ ameliorated the arsenic-induced testicular tissue injury observed by histopathological examination. In addition, TQ decreased the arsenic-induced expression of inducible nitric oxide synthase and caspase-3 in testicular tissue. It was concluded that TQ may represent a potential candidate to protect against arsenic-induced testicular injury. 

Keywords: Thymoquinone, arsenic, testes, rats.

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Authors: Trevor E. Davis, Isaac Cassar, Yi-Kai Lo, Wentai Liu

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This study models the use of transcutaneous electrical nerve stimulation on skin with a disk electrode in order to simulate tissue damage. The current density distribution above a disk electrode is known to be a dynamic and non-uniform quantity that is intensified at the edges of the disk. The non-uniformity is subject to change through using various electrode geometries or stimulation methods. One of these methods known as edge-retarded stimulation has shown to reduce this edge enhancement. Though progress has been made in modeling the behavior of a disk electrode, little has been done to test the validity of these models in simulating the actual heat transfer from the electrode. This simulation uses finite element software to couple the injection of current from a disk electrode to heat transfer described by the Pennesbioheat transfer equation. An example application of this model is studying an experimental form of stimulation, known as edge-retarded stimulation. The edge-retarded stimulation method will reduce the current density at the edges of the electrode. It is hypothesized that reducing the current density edge enhancement effect will, in turn, reduce temperature change and tissue damage at the edges of these electrodes. This study tests this hypothesis as a demonstration of the capabilities of this model. The edge-retarded stimulation proved to be safer after this simulation. It is shown that temperature change and the fraction of tissue necrosis is much greater in the square wave stimulation. These results bring implications for changes of procedures in transcutaneous electrical nerve stimulation and transcutaneous spinal cord stimulation as well.

Keywords: Bioheat transfer, Electrode, Neuroprosthetics, TENS, Transcutaneous stimulation.

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Authors: Amr A. Fouad, Waleed H. Albuali, Iyad Jresat

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The protective effect of hesperidin was investigated in rats exposed to liver injury induced by a single intraperitoneal injection of cyclophosphamide (CYP) at a dose of 150 mg kg-1. Hesperidin treatment (100 mg kg-1/day, orally) was applied for seven days, starting five days before CYP administration. Hesperidin significantly decreased the CYP-induced elevations of serum alanine aminotransferase, and hepatic malondialdehyde and myeloperoxidase activity, significantly prevented the depletion of hepatic glutathione peroxidase activity resulted from CYP administration. Also, hesperidin ameliorated the CYP-induced liver tissue injury observed by histopathological examination. In addition, hesperidin decreased the CYP-induced expression of inducible nitric oxide synthase, tumor necrosis factor-α, cyclooxygenase-2, Fas ligand, and caspase-9 in liver tissue. It was concluded that hesperidin may represent a potential candidate to protect against CYP-induced hepatotoxicity.

Keywords: Cyclophosphamide, hesperidin, liver, rats.

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Authors: M. A. Askari, M. A. Nazari, P. Perrier, Y. Payan

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Growth and remodeling of biological structures have gained lots of attention over the past decades. Determining the response of living tissues to mechanical loads is necessary for a wide range of developing fields such as prosthetics design or computerassisted surgical interventions. It is a well-known fact that biological structures are never stress-free, even when externally unloaded. The exact origin of these residual stresses is not clear, but theoretically, growth is one of the main sources. Extracting body organ’s shapes from medical imaging does not produce any information regarding the existing residual stresses in that organ. The simplest cause of such stresses is gravity since an organ grows under its influence from birth. Ignoring such residual stresses might cause erroneous results in numerical simulations. Accounting for residual stresses due to tissue growth can improve the accuracy of mechanical analysis results. This paper presents an original computational framework based on gradual growth to determine the residual stresses due to growth. To illustrate the method, we apply it to a finite element model of a healthy human face reconstructed from medical images. The distribution of residual stress in facial tissues is computed, which can overcome the effect of gravity and maintain tissues firmness. Our assumption is that tissue wrinkles caused by aging could be a consequence of decreasing residual stress and thus not counteracting gravity. Taking into account these stresses seems therefore extremely important in maxillofacial surgery. It would indeed help surgeons to estimate tissues changes after surgery.

Keywords: Finite element method, growth, residual stress, soft tissue.

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Authors: Gesine Hellwig

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Functional imaging procedures for the non-invasive assessment of tissue microcirculation are highly requested, but require a mathematical approach describing the trans- and intercapillary passage of tracer particles. Up to now, two theoretical, for the moment different concepts have been established for tracer kinetic modeling of contrast agent transport in tissues: pharmacokinetic compartment models, which are usually written as coupled differential equations, and the indicator dilution theory, which can be generalized in accordance with the theory of lineartime- invariant (LTI) systems by using a convolution approach. Based on mathematical considerations, it can be shown that also in the case of an open two-compartment model well-known from functional imaging, the concentration-time course in tissue is given by a convolution, which allows a separation of the arterial input function from a system function being the impulse response function, summarizing the available information on tissue microcirculation. Due to this reason, it is possible to integrate the open two-compartment model into the system-theoretic concept of indicator dilution theory (IDT) and thus results known from IDT remain valid for the compartment approach. According to the long number of applications of compartmental analysis, even for a more general context similar solutions of the so-called forward problem can already be found in the extensively available appropriate literature of the seventies and early eighties. Nevertheless, to this day, within the field of biomedical imaging – not from the mathematical point of view – there seems to be a trench between both approaches, which the author would like to get over by exemplary analysis of the well-known model.

Keywords: Functional imaging, Tracer kinetic modeling, LTIsystem, Indicator dilution theory / convolution approach, Two-Compartment model.

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Authors: Gintarė Sauliutė, Gintaras Svecevičius

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Landfill leachates contain a number of persistent pollutants, including heavy metals. They have the ability to spread in ecosystems and accumulate in fish which most of them are classified as top-consumers of trophic chains. Fish are freely swimming organisms; but perhaps, due to their species-specific ecological and behavioral properties, they often prefer the most suitable biotopes and therefore, did not avoid harmful substances or environments. That is why it is necessary to evaluate the persistent pollutant dispersion in hydroecosystem using fish tissue metal concentration. In hydroecosystems of hybrid type (e.g. river-pond-river) the distance from the pollution source could be a perfect indicator of such a kind of metal distribution. The studies were carried out in the Kairiai landfill neighboring hybrid-type ecosystem which is located 5 km east of the Šiauliai City. Fish tissue (gills, liver, and muscle) metal concentration measurements were performed on two types of ecologically-different fishes according to their feeding characteristics: benthophagous (Gibel carp, roach) and predatory (Northern pike, perch). A number of mathematical models (linear, non-linear, using log and other transformations) have been applied in order to identify the most satisfactorily description of the interdependence between fish tissue metal concentration and the distance from the pollution source. However, the only one log-multiple regression model revealed the pattern that the distance from the pollution source is closely and positively correlated with metal concentration in all predatory fish tissues studied (gills, liver, and muscle).

Keywords: Bioaccumulation in fish, heavy metals, hydroecosystem, landfill leachate, mathematical model.

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Authors: Fatahian Dehkordi RF., Parchami A.

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Design of experiments made for correlated the hormonal activity of steroids and their effect on the vaginal structure properties. Vaginal wall consists of distinct layers of cells which strongly differ regarding growth in answer to steroid hormones. The presence study carried out for functional evaluation and sustained replacement effect of the reproductive hormones on the vaginal morphometry in the rabbit. Rabbits were maintained control (Co) or ovariectomized (Ovz) and continuously treated with estradiol (Ovz E) and estradiol mixture with testosterone (Ovz ET). Relative to control rabbits with the distinct tissue distribution pattern, ovariectomized animals displayed declined epithelial thickness and atrophy of the muscularis layer that cause the vaginal wall thinning. Estradiol treatment of Orz rabbits inverted these changes to status of the preovariectomy. Histometric measurements in Ovz ET group completely including of thickness of the epithelial and muscular layers was greater in comparison to that of control animals.

Keywords: Vagina, Ovariectomy, Rabbit, Morphometry

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Authors: Lijiang Wang, Wei Wang, Yuhong Xu

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Lanthanide-doped upconversion nanoparticles which can convert near-infrared lights to visible lights have attracted growing interest because of their great potentials in fluorescence imaging. Upconversion fluorescence imaging technique with excitation in the near-infrared (NIR) region has been used for imaging of biological cells and tissues. However, improving the detection sensitivity and decreasing the absorption and scattering in biological tissues are as yet unresolved problems. In this present study, a novel NIR-reflected multispectral imaging system was developed for upconversion fluorescent imaging in small animals. Based on this system, we have obtained the high contrast images without the autofluorescence when biocompatible UCPs were injected near the body surface or deeply into the tissue. Furthermore, we have extracted respective spectra of the upconversion fluorescence and relatively quantify the fluorescence intensity with the multispectral analysis. To our knowledge, this is the first time to analyze and quantify the upconversion fluorescence in the small animal imaging.

Keywords: Multispectral imaging, near-infrared, upconversion fluorescence imaging, upconversion nanoparticles.

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Authors: Nour Ismail, Sahar El Kardawy, Bassant Badwy

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Radiofrequency (RF) lesioning of nerves have been commonly used to alleviate chronic pain, where RF current preventing transmission of pain signals through the nerve by heating the nerve causing the pain. There are some factors that affect the temperature distribution and the nerve lesion size, one of these factors is the inhomogeneities in the tissue medium. Our objective is to calculate the temperature distribution and the nerve lesion size in an inhomogeneous medium surrounding the RF electrode. A two 3-D finite element models are used to compare the temperature distribution in the homogeneous and inhomogeneous medium. Also the effect of temperature-dependent electric conductivity on maximum temperature and lesion size is observed. Results show that the presence of an inhomogeneous medium around the RF electrode has a valuable effect on the temperature distribution and lesion size. The dependency of electric conductivity on tissue temperature increased lesion size.

Keywords: Finite element model, nerve lesioning, pain relief, radiofrequency lesion.

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Authors: W. Kaowumpai, D. Koolpiruck, K. Viravaidya

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Female breast cancer is the second in frequency after cervical cancer. Surgery is the most common treatment for breast cancer, followed by chemotherapy as a treatment of choice. Although effective, it causes serious side effects. Controlled-release drug delivery is an alternative method to improve the efficacy and safety of the treatment. It can release the dosage of drug between the minimum effect concentration (MEC) and minimum toxic concentration (MTC) within tumor tissue and reduce the damage of normal tissue and the side effect. Because an in vivo experiment of this system can be time-consuming and labor-intensive, a mathematical model is desired to study the effects of important parameters before the experiments are performed. Here, we describe a 3D mathematical model to predict the release of doxorubicin from pluronic gel to treat human breast cancer. This model can, ultimately, be used to effectively design the in vivo experiments.

Keywords: Breast Cancer, Doxorubicin, Controlled ReleaseSystem, Diffusion and Convection Equation.

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Authors: J. Franková, V. Pivodová, F. Růžička, J. Ulrichová

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An implant elicits a biological response in the surrounding tissue which determines the acceptance and long-term function of the implant. Dental implants have become one of the main therapy methods in clinic after teeth lose. A successful implant is in contact with bone and soft tissue represent by fibroblasts. In our study we focused on the interaction between six different chemically and physically modified titanium implants (Tis-MALP, Tis-O, Tis- OA, Tis-OPAAE, Tis-OZ, Tis-OPAE) with alveolar fibroblasts as well as with five type of microorganisms (S. epidermis, S.mutans, S. gordonii, S. intermedius, C.albicans). The analysis of microorganism adhesion was determined by CFU (colony forming unite) and biofilm formation. The presence of α3β1 and vinculin expression on alveolar fibroblasts was demonstrated using phospho specific cell based ELISA (PACE). Alveolar fibroblasts have the highest expression of these proteins on Tis-OPAAE and Tis-OPAE. It corresponds with results from bacterial adhesion and biofilm formation and it was related to the lowest production of collagen I by alveolar fibroblasts on Tis-OPAAE titanium disc.

Keywords: titanium disc, alveolar fibroblasts, microorganisms, adhesion

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Authors: M. M. Bashandy, A. R. Ahmed, M. El-Gaffary, Sahar S. Abd El-Rahman

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This study evaluated the acute toxicity and tissue distribution of intravenously administered gold nanoparticles (AuNPs) in male rabbits. Rabbits were exposed to single dose of AuNPs (300 μg/ kg). Toxic effects were assessed via general behavior, hematological parameters, serum biochemical parameters, and histopathological examination of various rabbits’ organs. Inductively coupled plasma–mass spectrometry (ICP-MS) was used to determine gold concentrations in tissue samples collected at predetermined time intervals. After one week, AuNPs exerted no obvious acute toxicity in rabbits. However, inflammatory reactions were observed in liver, lungs and kidneys accompanied with mild absolute neutrophilia and significant monocytosis. The highest gold levels were found in the spleen and liver followed by lungs, and kidneys. These results indicated that AuNPs could be distributed extensively to various tissues in the body, but primarily in the spleen and liver.

Keywords: Gold nanoparticles, toxicity, pathology, hematology, liver function, kidney function.

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Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H₂O and CO₂. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: Poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblasts, rat vascular smooth muscle cells, human stem cells.

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Authors: Elhamalsadat Ghaffari, Moghan Amirhosseinian, Amir Khaleghipour

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Multifunctional bioactive glasses (BGs) are designed with a focus on the provision of bactericidal and biological properties desired for angiogenesis, osteogenesis, and ultimately potential applications in bone tissue engineering. To achieve these, six sol-gel copper/magnesium substituted derivatives of 58S-BG, i.e. a mol% series of 60SiO₂-4P₂O₅-5CuO-(31-x) CaO/xMgO (where x=0, 1, 3, 5, 8, and 10), were synthesized. Afterwards, the effect of MgO/CaO substitution on the in vitro formation of nano-hydroxyapatite (HA), osteoblast-like cell responses and BGs antibacterial performance were studied. During the BGs synthesis, the elimination of nitrates was achieved at 700 °C that prevented the BGs crystallization and stabilized the obtained dried gels. The structural and morphological evaluations were performed with X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). These characterizations revealed that Cu-substituted 58S-BG consisting of 5 mol% MgO (BG-5/5) slightly had retarded the formation of HA. In addition, Cu-substituted 58S-BGs consisting 8 mol% and 10 mol% MgO (BG-5/8 and BG-5/10) displayed lower bioactivity probably due to the lower ion release rate of Ca–Si into the simulated body fluid (SBF). The determination of 3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and alkaline phosphate (ALP) activities proved that the highest values of both differentiation and proliferation of MC3T3-E1 cells can be obtained from a 5 mol% MgO substituted BG, while the over addition of MgO (8 mol% and 10 mol%) decreased the bioactivity. Furthermore, these novel Cu/Mg-substituted 58S-BGs displayed antibacterial effect against methicillin-resistant Staphylococcus aureus bacteria. Taken together, the results suggest the equally-substituted BG-5/5 (i.e. the one consists of 5 mol% of both CuO and MgO) as a promising candidate for bone tissue engineering, among all newly designed BGs in this work, owing to its desirable cell proliferation, ALP activity and antibacterial properties.

Keywords: Apatite, bioactivity, biomedical applications sol-gel processes.

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Authors: Chih-Wei Chao, Jiashing Yu

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To mimic the natural circumstances of cell growth in an organism, we present three-dimensional (3D) scaffolds fabricated by microfluidics for cultivation. This work investigates the cellular behaviors of rat cardiomyocytes in gelatin 3D scaffolds compared to those on 2D control, such as proliferation, viability and morphology. We found that the scaffolds may induce skeletal differentiation of H9c2 cells.

Keywords: Microfluidic device, H9c2, tissue engineering, 3D scaffolds.

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Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

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Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA₄₄₀₄. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: Recombinant tissue plasminogen activator, plant cell comportment, leader signals, transgenic tobacco.

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Authors: Rafael Muñoz, Juan Melchor, Alicia Valera, Laura Peralta, Guillermo Rus

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A torsional piezoelectric ultrasonic transducer design is proposed to measure shear moduli in soft tissue with direct access availability, using shear wave elastography technique. The measurement of shear moduli of tissues is a challenging problem, mainly derived from a) the difficulty of isolating a pure shear wave, given the interference of multiple waves of different types (P, S, even guided) emitted by the transducers and reflected in geometric boundaries, and b) the highly attenuating nature of soft tissular materials. An immediate application, overcoming these drawbacks, is the measurement of changes in cervix stiffness to estimate the gestational age at delivery. The design has been optimized using a finite element model (FEM) and a semi-analytical estimator of the probability of detection (POD) to determine a suitable geometry, materials and generated waves. The technique is based on the time of flight measurement between emitter and receiver, to infer shear wave velocity. Current research is centered in prototype testing and validation. The geometric optimization of the transducer was able to annihilate the compressional wave emission, generating a quite pure shear torsional wave. Currently, mechanical and electromagnetic coupling between emitter and receiver signals are being the research focus. Conclusions: the design overcomes the main described problems. The almost pure shear torsional wave along with the short time of flight avoids the possibility of multiple wave interference. This short propagation distance reduce the effect of attenuation, and allow the emission of very low energies assuring a good biological security for human use.

Keywords: Cervix ripening, preterm birth, shear modulus, shear wave elastography, soft tissue, torsional wave.

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Authors: Cheng-Hong Yang, Tsung-Mu Shih, Li-Yeh Chuang

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Serial Analysis of Gene Expression is a powerful quantification technique for generating cell or tissue gene expression data. The profile of the gene expression of cell or tissue in several different states is difficult for biologists to analyze because of the large number of genes typically involved. However, feature selection in machine learning can successfully reduce this problem. The method allows reducing the features (genes) in specific SAGE data, and determines only relevant genes. In this study, we used a genetic algorithm to implement feature selection, and evaluate the classification accuracy of the selected features with the K-nearest neighbor method. In order to validate the proposed method, we used two SAGE data sets for testing. The results of this study conclusively prove that the number of features of the original SAGE data set can be significantly reduced and higher classification accuracy can be achieved.

Keywords: Serial Analysis of Gene Expression, Feature selection, Genetic Algorithm, K-nearest neighbor method.

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Authors: Mamoun F. Al-Mistarihi

Abstract:

We have previously introduced an ultrasonic imaging approach that combines harmonic-sensitive pulse sequences with a post-beamforming quadratic kernel derived from a second-order Volterra filter (SOVF). This approach is designed to produce images with high sensitivity to nonlinear oscillations from microbubble ultrasound contrast agents (UCA) while maintaining high levels of noise rejection. In this paper, a two-step algorithm for computing the coefficients of the quadratic kernel leading to reduction of tissue component introduced by motion, maximizing the noise rejection and increases the specificity while optimizing the sensitivity to the UCA is presented. In the first step, quadratic kernels from individual singular modes of the PI data matrix are compared in terms of their ability of maximize the contrast to tissue ratio (CTR). In the second step, quadratic kernels resulting in the highest CTR values are convolved. The imaging results indicate that a signal processing approach to this clinical challenge is feasible.

Keywords: Volterra Filter, Pulse Inversion, Ultrasonic Imaging, Contrast Agent.

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Authors: Saowaluk Sikiwat, Mayuree Pumipaiboon, Sutee Kaewsangiem, Mayuva Areekijseree

Abstract:

The problem on the conservation programme of the Royal Thai Navy Sea Turtle Nursery, Phang-nga Province, Thailand is high mortality rate of juvenile green sea turtle (Cheloniamydas) on nursing period. So, during May to October 2012, postmortem examinations of juvenile green sea turtle were performed to determine the causes of dead. Fresh tissues of postmortem of 15 juvenile green sea turtles (1-3 months old) were investigated using paraffin section technique. The results showed normal ultrastructure of all tissue organs. These instances reviewed the health and stability of the environments in which juvenile green sea turtles live and concern for the survival rate. The present article also provides guidance for a review of the biology, guidelines for appropriate postmortem tissue, normal histology and sampling collection and procedures. The data also provides information for conservation of this endangered species in term of acknowledging and encouraging people to protect the animals and their habitats in nature.

Keywords: Green sea turtles (Cheloniamydas), histology, juvenile sea turtles

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Authors: L. V. Ignatova, Y. V. Brazhnikova, T. D. Mukasheva, R. Zh. Berzhanova, A. A. Omirbekova

Abstract:

Endophytic microorganisms are presented in plants of different families growing in the foothills and piedmont plains of Trans-Ili Alatau. It was found that the maximum number of endophytic micromycetes is typical to the Fabaceae family. The number of microscopic fungi in the roots reached (145.9±5.9)×10³ CFU/g of plant tissue; yeasts - (79.8±3.5)×10² CFU/g of plant tissue. Basically, endophytic microscopic fungi are typical for underground parts of plants. In contrast, yeasts more infected aboveground parts of plants. Small amount of micromycetes is typical to inflorescence and fruits. Antagonistic activity of selected micromycetes against Fusarium graminearum, Cladosporium sp., Phytophtora infestans and Botrytis cinerea phytopathogens was detected. Strains with a broad, narrow and limited range of action were identified. For further investigations Rh2 and T7 strains were selected, they are characterized by a broad spectrum of fungicidal activity and they formed the large inhibition zones against phytopathogens. Active antagonists are attributed to the Rhodotorula mucilaginosa and Beauveria bassiana species.

Keywords: Endophytic micromycetes, fungicidal activity, prevalence.

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Authors: Mohammad Shafigh, Nasser Fatouraee, Amirsaied Seddighi

Abstract:

Mechanical properties of cerebral arteries are, due to their relationship with cerebrovascular diseases, of clinical worth. To acquire these properties, eight samples were obtained from middle cerebral arteries of human cadavers, whose death were not due to injuries or diseases of cerebral vessels, and tested within twelve hours after resection, by a precise biaxial tensile test device specially developed for the present study considering the dimensions, sensitivity and anisotropic nature of samples. The resulting stress-stretch curve was plotted and subsequently fitted to a hyperelastic three-parameter Fung model. It was found that the arteries were noticeably stiffer in circumferential than in axial direction. It was also demonstrated that the use of multi-parameter hyperelastic constitutive models is useful for mathematical description of behavior of cerebral vessel tissue. The reported material properties are a proper reference for numerical modeling of cerebral arteries and computational analysis of healthy or diseased intracranial arteries.

Keywords: Anisotropic Tissue, Cerebral Blood Vessels, Fung Model, Nonlinear Material, Plain Stress.

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Authors: Daniel Olmos-Liceaga

Abstract:

Ventricular fibrillation is a very important health problem as is the cause of most of the sudden deaths in the world. Waves of electrical activity are sent by the SA node, propagate through the cardiac tissue and activate the mechanisms of cell contraction, and therefore are responsible to pump blood to the body harmonically. A spiral wave is an abnormal auto sustainable wave that is responsible of certain types of arrhythmias. When these waves break up, give rise to the fibrillation regime, in which there is a complete loss in the coordination of the contraction of the heart muscle. Interaction of spiral waves and obstacles is also of great importance as it is believed that the attachment of a spiral wave to an obstacle can provide with a transition of two different arrhythmias. An obstacle can be partially excitable or non excitable. In this talk, we present a numerical study of the interaction of meandering spiral waves with partially and non excitable obstacles and focus on the problem where the obstacle plays a fundamental role in the switch between different spiral regimes, which represent different arrhythmic regimes. Particularly, we study the phenomenon of destabilization of spiral waves due to the presence of obstacles, a phenomenon not completely understood (This work will appear as a Chapter in a Book named Cardiac Arrhytmias by INTECH under the name "Spiral Waves, Obstacles and Cardiac Arrhythmias", ISBN 979-953-307-050-5.).

Keywords: Arrhythmias, Cardiac tissue, Obstacles, Spiral waves

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Authors: M.S.Nourbakhsh, M.E.khosroshahi

Abstract:

Gold coated silica core nanoparticles have an optical response dictated by the plasmon resonance. The wavelength at which the resonance occurs depends on the core and shell sizes, allowing nanoshells to be tailored for particular applications. The purposes of this study was to synthesize and use different concentration of gold nanoshells as exogenous material for skin tissue soldering and also to examine the effect of laser soldering parameters on the properties of repaired skin. Two mixtures of albumin solder and different concentration of gold nanoshells were prepared. A full thickness incision of 2×20 mm2 was made on the surface and after addition of mixtures it was irradiated by an 810nm diode laser at different power densities. The changes of tensile strength σt due to temperature rise, number of scan (Ns), and scan velocity (Vs) were investigated. The results showed at constant laser power density (I), σt of repaired incisions increases by increasing the concentration of gold nanoshells, Ns and decreasing Vs. It is therefore important to consider the trade off between the scan velocity and the surface temperature for achieving an optimum operating condition. In our case this corresponds to σt =1610 gr/cm2 at I~ 60 Wcm-2, T ~ 65ºC, Ns =10 and Vs=0.2mms-1.

Keywords: Tissue soldering, Diode laser, Gold Nanoshells, Tensile strength

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