Abstract: Ischemic events can culminate in acute myocardial infarction with irreversible cardiac lesions that cannot be restored due to the limited regenerative capacity of the heart. Tissue engineering proposes therapeutic alternatives by using biomaterials to resemble the native extracellular medium combined with healthy and functional cells. This research focused on developing a natural thermosensitive hydrogel, its physical-chemical characterization and in vitro biocompatibility determination. Hydrogels’ morphological characterization was carried out through scanning electron microscopy and its chemical characterization by employing Infrared Spectroscopy technic. In addition, the biocompatibility was determined using fetal human ventricular cardiomyocytes cell line RL-14 and the MTT cytotoxicity test according to the ISO 10993-5 standard. Four biocompatible and thermosensitive hydrogels were obtained with a three-dimensional internal structure and two gelation times. The results show the potential of the hydrogel to increase the cell survival rate to the cardiac cell therapies under investigation and lay the foundations to continue with its characterization and biological evaluation both in vitro and in vivo models.
Abstract: Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.
Abstract: In this study, the mechanical properties of alginate hydrogel material for self-standing 3D scaffold architecture with proper shape fidelity are investigated. In-lab built 3D bio-printer extrusion-based technology is utilized to fabricate 3D alginate scaffold constructs. The pressure, needle speed and stage speed are varied using a computer-controlled system. The experimental result indicates that the concentration of alginate solution, calcium chloride (CaCl2) cross-linking concentration and cross-linking ratios lead to the formation of alginate hydrogel with various gelation states. Besides, the gelling conditions, such as cross-linking reaction time and temperature also have a significant effect on the mechanical properties of alginate hydrogel. Various experimental tests such as the material gelation, the material spreading and the printability test for filament collapse as well as the swelling test were conducted to evaluate the fabricated 3D scaffold constructs. The result indicates that the fabricated 3D scaffold from composition of 3.5% wt alginate solution, that is prepared in DI water and 1% wt CaCl2 solution with cross-linking ratios of 7:3 show good printability and sustain good shape fidelity for more than 20 days, compared to alginate hydrogel that is prepared in a phosphate buffered saline (PBS). The fabricated self-standing 3D scaffold constructs measured 30 mm × 30 mm and consisted of 4 layers (n = 4) show good pore geometry and clear grid structure after printing. In addition, the percentage change of swelling degree exhibits high swelling capability with respect to time. The swelling test shows that the geometry of 3D alginate-scaffold construct and of the macro-pore are rarely changed, which indicates the capability of holding the shape fidelity during the incubation period. This study demonstrated that the mechanical and physical properties of alginate hydrogel could be tuned for a 3D bio-printing extrusion-based system to fabricate self-standing 3D scaffold soft structures. This 3D bioengineered scaffold provides a natural microenvironment present in the extracellular matrix of the tissue, which could be seeded with the biological cells to generate the desired 3D live tissue model for in vitro and in vivo tissue engineering applications.
Abstract: Magnetically controlled growing rods (MCGRs) have been used to stabilise and correct spinal curvature in children to support non-invasive scoliosis adjustment. Although the encapsulated driving components are intended to be isolated from body fluid contact, in vivo corrosion was observed on these components due to sealing mechanism damage. Consequently, a corrosion circuit is created with the body fluids, resulting in malfunction of the lengthening mechanism. Particularly, the chloride ions in blood plasma or cerebrospinal fluid (CSF) may corrode the MCGR alloys, possibly resulting in metal ion release in long-term use. However, there is no data available on the corrosion resistance of spinal implant alloys in CSF. In this study, an in vitro immersion configuration was designed to simulate in vivo corrosion of 440C SS-Ti6Al4V couples. The 440C stainless steel (SS) was heat-treated to investigate the effect of tempering temperature on intergranular corrosion (IGC), while crevice and galvanic corrosion were studied by limiting the clearance of dissimilar couples. Tests were carried out in a neutral artificial cerebrospinal fluid (ACSF) and phosphate-buffered saline (PBS) under aeration and deaeration for 2 months. The composition of the passive films and metal ion release were analysed. The effect of galvanic coupling, pH, dissolved oxygen and anion species on corrosion rates and corrosion mechanisms are discussed based on quantitative and qualitative measurements. The results suggest that ACSF is more aggressive than PBS due to the combination of aggressive chlorides and sulphate anions, while phosphate in PBS acts as an inhibitor to delay corrosion. The presence of Vivianite on the SS surface in PBS lowered the corrosion rate (CR) more than 5 times for aeration and nearly 2 times for deaeration, compared with ACSF. The CR of 440C is dependent on passive film properties varied by tempering temperature and anion species. Although the CR of Ti6Al4V is insignificant, it tends to release more Ti ions in deaerated ACSF than under aeration, about 6 µg/L. It seems the crevice-like design has more effect on macroscopic corrosion than combining the dissimilar couple, whereas IGC is dominantly observed on sensitized microstructure.
Abstract: Alzheimer’s disease (AD) (a progressive neurodegenerative disorder) is mostly predominant cause of dementia in the elderly. Prolonging the function of acetylcholine by inhibiting both acetylcholinesterase and butyrylcholinesterase is most effective treatment therapy of AD. Traditionally Pterocarpus santalinus L. is widely known for its medicinal use. In this study, in vitro acetylcholinesterase inhibitory activity was investigated and methanolic extract of the plant showed significant activity. To confirm this activity (in vivo), learning and memory enhancing effects were tested in mice. For the test, memory impairment was induced by scopolamine (cholinergic muscarinic receptor antagonist). Anti-amnesic effect of the extract was investigated by the passive avoidance task in mice. The study also includes brain acetylcholinesterase activity. Results proved that scopolamine induced cognitive dysfunction was significantly decreased by administration of the extract solution, in the passive avoidance task and inhibited brain acetylcholinesterase activity. These results suggest that bark extract of Pterocarpus santalinus can be better option for further studies on AD via their acetylcholinesterase inhibitory actions.
Abstract: A hydrogel from cellulose acetate cross linked with ethylenediaminetetraacetic dianhydride (HAC-EDTA) was synthesized by our research group, and submitted to characterization and biological tests. Cytocompatibility analysis was performed by confocal microscopy using human adipocyte derived stem cells (ASCs). The FTIR analysis showed characteristic bands of cellulose acetate and hydroxyl groups and the tensile tests evidence that HAC-EDTA present a Young’s modulus of 643.7 MPa. The confocal analysis revealed that there was cell growth at the surface of HAC-EDTA. After one day of culture the cells presented spherical morphology, which may be caused by stress of the sequestration of Ca2+ and Mg2+ ions at the cell medium by HAC-EDTA, as demonstrated by ICP-MS. However, after seven days and 14 days of culture, the cells present fibroblastoid morphology, phenotype expected by this cellular type. The results give efforts to indicate this new material as a potential biomaterial for tissue engineering, in the future in vivo approach.
Abstract: X-ray Fluorescence Molecular Imaging (XFMI) holds great promise as a low-cost molecular imaging modality for biomedical applications with high chemical sensitivity. However, for in vivo biomedical applications, a key technical bottleneck is the relatively low chemical sensitivity of XFMI, especially at a reasonably low radiation dose. In laboratory x-ray source based XFMI, one of the main factors that limits the chemical sensitivity of XFMI is the scattered x-rays. We will present our latest findings on improving the chemical sensitivity of XFMI using excitation beam spectrum optimization. XFMI imaging experiments on two mouse-sized phantoms were conducted at three different excitation beam spectra. Our results show that the minimum detectable concentration (MDC) of iodine can be readily increased by five times via excitation spectrum optimization. Findings from this investigation could find use for in vivo pre-clinical small-animal XFMI in the future.
Abstract: Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.
Abstract: Mechanical index (MI) is used for quantifying acoustic cavitation and the relationship between acoustic pressure and the frequency. In this study, modeling of the MI was applied to provide treatment protocol and to understand the effective physical processes on reproducibility of stem cells. The acoustic pressure and MI equations are modeled and solved to estimate optimal MI for 28, 40, 150 kHz and 1 MHz frequencies. Radial and axial acoustic pressure distribution was extracted. To validate the results of the modeling, the acoustic pressure in the water and near field depth was measured by a piston hydrophone. Results of modeling and experiments show that the model is consistent well to experimental results with 0.91 and 0.90 correlation of coefficient (p
Abstract: The propose of this study was to investigate in vitro thrombolytic activity of Zingiber cassumunar Roxb. and Prasaplai, a Thai herbal formulation of Z. cassumunar Roxb. Herbs were extracted with boiling water and concentrated by lyophilization. To observe their thrombolytic potential, an in vitro clot lysis method was applied where streptokinase and sterile distilled water were used as positive and negative controls, respectively. Crude aqueous extracts from Z. cassumunar Roxb. and Prasaplai formula showed significant thrombolytic activity by clot lysis of 17.90% and 25.21%, respectively, compared to the negative control water (5.16%) while the standard streptokinase revealed 64.78% clot lysis. These findings suggest that Z. cassumunar Roxb. exhibits moderate thrombolytic activity and cloud play an important role in the thrombolytic properties of Prasaplai formula. However, further study should be done to observe in vivo clot dissolving potential and to isolate active component(s) of these extracts.
Abstract: Nowadays, nanoparticles are being used in pharmacological studies for their exclusive properties such as small size, more surface area, biocompatibility and enhanced solubility. In view of this, the present study aimed to evaluate the antihyperglycemic potential of biologically synthesized silver nanoparticles (BSSNPs) and Gymnema sylvestre (GS) extract. The SEM and SEM analysis divulges that the BSSNPs were spherical in shape. EDAX spectrum exhibits peaks for the presence of silver, carbon, and oxygen atoms in the range of 1.0-3.1 keV. FT-IR reveals the binding properties of active bio-constituents responsible for capping and stabilizing BSSNPs. The results showed increased blood glucose, huge loss in body weight and downturn in plasma insulin. The GS extract (200 mg/kg, 400 mg/kg), BSSNPs (100 mg/kg, 200 mg/kg) and metformin 50 mg/kg were administered to the diabetic rats. BSSNPs at a dose level of 200 mg/kg (b.wt.p.o.) showed significant inhibition of (p
Abstract: Angiogenesis, a formation of new blood vessels from a pre-existing vasculature, plays an important role in pathologic processes such as the growth and metastasis of tumours. Tumours cannot grow beyond a few millimetres without blood supply from the newly formed blood vessels from the host tissue, a process called tumour-induced angiogenesis. The successful research of antiangiogenic treatment of cancer has focused on nutraceuticals with angiogenesis-modulating properties. Berberine, as a major active component of the bark of Phellodendron amurense Rupr., has shown antitumour activity by intervening into different steps of carcinogenesis. The influence of ethanolic extract of Phellodendron amurese bark on the angiogenesis was tested in vivo on chick chorioallantoic membrane (CAM). The irritancy of the CAM after the application of the crude bark extract dissolved in normal saline (10 mg/mL) was investigated on embryonic day 7. No significant signs of the irritancy, such as vasoconstriction, hyperaemia, haemorrhage or coagulation were observed which indicates the harmless character of the extract. A significant reduction in vessel sprouting and higher percentage of avascular zone was observed in the case of CAM treated with the extract in comparison with non-treated CAM (control), which is a proof of the antiangiogenic potential of the extract. These results could contribute to the development of novel drugs for the treatment of cancer or other diseases, in which angiogenesis plays a significant role.
Abstract: Human soft tissue is loaded and deformed by any
activity, an effect known as a stress-strain relationship, and is often
described by a load and tissue elongation curve. Several advances
have been made in the fields of biology and mechanics of soft human
tissue. However, there is limited information available on in vivo
tissue mechanical characteristics and behavior. Confident mechanical
properties of human soft tissue cannot be extrapolated from e.g.
animal testing. Thus, there is need for non invasive methods to
analyze mechanical characteristics of soft human tissue. In the present
study, the internal mechanical conditions of the lower limb, which
is subject to an external load, is studied by use of the finite element
method. A detailed finite element model of the lower limb is made
possible by use of MRI scans. Skin, fat, bones, fascia and muscles
are represented separately and the material properties for them are
obtained from literature. Previous studies have been shown to address
macroscopic deformation features, e.g. indentation depth, to a large
extent. However, the detail in which the internal anatomical features
have been modeled does not reveal the critical internal strains that
may induce hypoxia and/or eventual tissue damage. The results of the
present study reveals that lumped material models, i.e. averaging of
the material properties for the different constituents, does not capture
regions of critical strains in contrast to more detailed models.
Abstract: Niosomes were formulated with an aim of enhancing the oral bioavailability of losartan potassium and formulated in different molar ratios of surfactant, cholesterol and dicetyl phosphate. The formulated niosomes were found in range of 54.98 µm to 107.85 µm in size. Formulations with 1:1 ratio of surfactant and cholesterol have shown maximum entrapment efficiencies. Niosomes with sorbitan monostearate showed maximum drug release and zero order release kinetics, at the end of 24 hours. The in vivo study has shown the significant enhancement in oral bioavailability of losartan potassium in rats, after a dose of 10 mg/kg. The average relative bioavailability in relation with pure drug solution was found 2.56, indicates more than two fold increase in oral bioavailability. A significant increment in MRT reflects the release retarding ability of the vesicles. In conclusion, niosomes could be a promising delivery of losartan potassium with improved oral bioavailability and prolonged release profiles.
Abstract: Many herbal medicinal products are considered
potential anti-hypercholesterolemic agents with encouraging safety
profiles, however only a limited amount of clinical research exists to
support their efficacy. The present study was designed to compare the
antihypercholesterolemic and antioxidant activities of the crude
ethanolic extracts of Citrus reticulata fruit peel, Zingiber officinale
rhizome and Sesamum indicum seeds. Forty-five rats were used throughout the experiment which are
extended for four weeks. These were divided into nine groups, five
rats per each group as follows; group 1 was the normal control group
(rats only fed standard normal rat diet), group 2 was the
hypercholesterolemic control group (rats fed only
hypercholesterolemic diet which contained 1% cholesterol plus 10%
saturated animal fat added to the normal rat diet), groups 3 and 4
were fed hypercholesterolemic diet in addition to Citrus reticulata
ethanolic extract at doses of (250mg/kg (group 3) and 500mg/kg
(group 4)) administered daily via oral route, groups 5 and 6 were
given hypercholesterolemic diet in addition to Zingiber officinale
ethanolic extract at doses of (250mg/kg (group 5) and 500mg/kg
(group 6)) daily through oral route, groups 7 and 8 fed on
hypercholesterolemic diet in addition to Sesamum indicum ethanolic
extract at doses of (250mg/kg (group 7) and 500mg/kg (group 8))
daily orally; and group 9 rats were given hypercholesterolemic diet in
addition to atorvastatin (0.18mg/kg) daily via oral route as a standard
reference antihypercholesterolemic drug. Blood samples from all
groups were drawn from the retro-orbital venous plexus four weeks
following treatment after overnight fasting and the lipid profile (total
cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low
density lipoprotein-cholesterol (LDL-C) and triglyceride levels) were
measured and the risk ratio (TC/HDL-C) was assessed. The
antioxidant activity of the three plants extracts was determined using
DPPH free-radical antioxidant assay. Results of in vivo and in vitro
antihypercholesterolemic and antioxidant assay respectively, revealed
that the three extracts possess comparable antioxidant and
antihypercholesterolemic activities.
Abstract: This report presents an alternative technique of
application of contrast agent in vivo, i.e. before sampling. By this
new method the electron micrograph of tissue sections have an
acceptable contrast compared to other methods and present no artifact
of precipitation on sections. Another advantage is that a small amount
of contrast is needed to get a good result given that most of them are
expensive and extremely toxic.
Abstract: SARA is a common and serious metabolic disorder in
early lactation in dairy cattle and in finishing beef cattle, caused by
diets with high inclusion of cereal grain. This experiment was
performed to determine the efficacy of Megasphaera elsdenii, a
major lactate-utilizing bacterium in prevention/treatment of SARA in
vivo. In vivo experimentation, it was used eight ruminally cannulated
rams and it was applied the rapid adaptation with the mixture of grain
based on wheat (80% wheat, 20% barley) and barley (80% barley,
20% wheat). During the systematic adaptation, it was followed the
probability of SARA formation by being measured the rumen pH
with two hours intervals after and before feeding. After being
evaluated the data, it was determined the ruminal pH ranged from
5.2-5.6 on the condition of feeding with 60 percentage of grain
mixture based on barley and wheat, that assured the definite form of
subacute acidosis. In four days SARA period, M. elsdenii (1010 cfu
ml-1) was inoculated during the first two days. During the SARA
period, it was observed the decrease of feed intake with M. elsdenii
inoculation. Inoculation of M. elsdenii was caused to differentiation
of rumen pH (P
Abstract: Doxorubicin (DOX) is an anthracycline drug used to treat many cancer diseases. Similarly to other cytostatic drugs, DOX has serious side effects; the biggest obstacle is the cardiotoxicity. With the aim of lowering the negative side effects and to target the DOX into the tumor tissue, the different nanoparticles (NPs) are studied. The aim of this work was to synthetized different NPs and conjugated them with DOX and determine the binding capacity of the NPs. For this experiment, carbon nanotubes (CNTs), graphene oxide (GO), fullerene (FUL) and liposomes (LIP) were used. The highest binding capacity was observed in GO (85%). Subsequently the toxicity of NPs and NPs-DOX conjugates was analyzed in in vivo system (chicken embryos). Some NPs (GO) can increase the toxicity of DOX, whereas other NPs (LIP, CNTs) decrease DOX toxicity.
Abstract: In the immunologic sense, clinical infection is a state
of failure of the immune system to combat the pathogenic weapon of
the bacteria invading the host. A motile gram negative vibroid
organism associated with marked mono and poly nuclear cell
responses was traced during the examination of a clinical material
from an infected common carp Cyprinus carpio. On primary plate
culture, growth was shown to be pure, dense population of an
Aeromonas-like colony morphotype. The pure isolate was found to
be; Aerobic, facultatively anaerobic, non-halophilic, grew at 0C, and
37C, oxidase positive utilizes glucose through fermentative pathway,
resist 0/129 and novobiocin, produces alanine and lysine
decarboxylases but non-producing ornithine dehydrolases. Tests for
the in vitro determinants of pathogenicity has shown to be; Betahaemolytic
onto blood agar, gelatinase, casienase and amylase
producer. Three in vivo determinants of pathogenicity were tested as,
the lethal dose fifty, the pathogenesis and pathogenicity. It was
evident that 0.1 milliliter of the causal bacterial cell suspension of a
density 1 x 107 CFU/ml injected intramuscularly into an average of
100gms fish toke five days incubation period, then at the day six
morbidity and mortality were initiated. LD50 was recorded at the day
12 post-infection. Use of an LD50 doses to study the pathogenicity,
reveals mononuclear and polynuclear cell responses, on examining
the stained direct films of the clinical materials from the
experimentally infected fish. Re-isolation tests confirm that the reisolant
is same. The course of the infection in natural case was shown
manifestation of; skin ulceration, haemorrhage and descaling. On
evisceration, the internal organs were shown; congestion in the
intestines, spleen and, air sacs. The induced infection showed a
milder form of these manifestations. The grading of the virulence of
this organism was virulent causing chronic course of infections as
indicated from the pathogenesis and pathogenicity studies. Thus the
infectious bacteria were consistent with Aeromonas hydrophila, and
the infection was chronic.
Abstract: Enterococci are important inhabitants of the animal
intestine and are widely used in probiotic products. A probiotic strain
is expected to possess several desirable properties in order to exert
beneficial effects. Therefore, the objective of this study was to
isolate, characterize and identify Enterococcus sp. from chicken cecal
and fecal samples to determine potential probiotic properties.
Enterococci were isolated from chicken ceca and feces of thirty three
clinically healthy chickens from a local farm. In vitro studies were
performed to assess antibacterial activity of the isolated LAB (using
agar well diffusion and cell free supernatant broth technique against
Salmonella enterica serotype Enteritidis), survival in acidic
conditions, resistance to bile salts, and their survival during simulated
gastric juice conditions at pH 2.5. Isolates were identified by
biochemical carbohydrate fermentation patterns using an API 50
CHL kit and API ZYM kits and by sequenced 16S rDNA. An isolate
belonging to E. faecium species exhibited inhibitory effect against S.
enteritidis. This isolate producing a clear zone as large as 10.30 mm
or greater and was able to grow in the coculture medium and at the
same time, inhibited the growth S. enteritidis. In addition, E. faecium
exhibited significant resistance under highly acidic conditions at pH
2.5 for 8 h and survived well in bile salt at 0.2% for 24 h and showing
ability to survive in the presence of simulated gastric juice at pH 2.5.
Based on these results, E. faecium isolate fulfills some of the criteria
to be considered as a probiotic strain and therefore, could be used as a
feed additive with good potential for controlling S. Enteritidis in
chickens. However, in vivo studies are needed to determine the safety
of the strain.