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Mechano-pharmacological testing of L-Type Ca²⁺ channel modulators via human vascular celldrum model
(2020)
Background/Aims: This study aimed to establish a precise and well-defined working model, assessing pharmaceutical effects on vascular smooth muscle cell monolayer in-vitro. It describes various analysis techniques to determine the most suitable to measure the biomechanical impact of vasoactive agents by using CellDrum technology. Methods: The so-called CellDrum technology was applied to analyse the biomechanical properties of confluent human aorta muscle cells (haSMC) in monolayer. The cell generated tensions deviations in the range of a few N/m² are evaluated by the CellDrum technology. This study focuses on the dilative and contractive effects of L-type Ca²⁺ channel agonists and antagonists, respectively. We analyzed the effects of Bay K8644, nifedipine and verapamil. Three different measurement modes were developed and applied to determine the most appropriate analysis technique for the study purpose. These three operation modes are called, particular time mode" (PTM), "long term mode" (LTM) and "real-time mode" (RTM). Results: It was possible to quantify the biomechanical response of haSMCs to the addition of vasoactive agents using CellDrum technology. Due to the supplementation of 100nM Bay K8644, the tension increased approximately 10.6% from initial tension maximum, whereas, the treatment with nifedipine and verapamil caused a significant decrease in cellular tension: 10nM nifedipine decreased the biomechanical stress around 6,5% and 50nM verapamil by 2,8%, compared to the initial tension maximum. Additionally, all tested measurement modes provide similar results while focusing on different analysis parameters. Conclusion: The CellDrum technology allows highly sensitive biomechanical stress measurements of cultured haSMC monolayers. The mechanical stress responses evoked by the application of vasoactive calcium channel modulators were quantified functionally (N/m²). All tested operation modes resulted in equal findings, whereas each mode features operation-related data analysis.
The carbonized rice husk (CRH) was evaluated for its wound healing activity in rats using excision models. In this study, the influences of CRH on wound healing in rat skin in vivo and cellular behavior of human dermal fibroblasts in vitro were investigated. The obtained results showed that the CRH treatment promoted wound epithelization in rats and exhibited moderate inhibition of cell proliferation in vitro. CRH with lanolin oil treated wounds were found to epithelize faster as compared to controls.
Recently, the SHARP Corporation, Japan, has developed the world’s first "Plasma Cluster Ions (PCI)" air purification technology using plasma discharge to generate cluster ions. The new plasma cluster device releases positive and negative ions into the air, which are able to decompose and deactivate harmful airborne substances by chemical reactions. Because cluster ions consist of positive and negative ions that normally exist in the natural world, they are completely harmless and safe to humans. The amount of ozone generated by cluster ions is less than 0.01 ppm, which is significantly less than the 0.05-ppm standard for industrial operations and consumer electronics. This amount, thus, has no harming effects whatsoever on the human body. But particular properties and chemical processes in PCI treatment are still under study. It has been shown that PCI in most cases show strongly pronounced irreversible killing effects in respect of airborne microflora due to free-radical induced reactions and can be considered as a potent technology to disinfect both home, medical and industrial appliances.
Bacterial lipopolysaccharides (endotoxins) show strong biological effects at very low concentrations in human beings and many animals when entering the blood stream. These include affecting structure and function of organs and cells, changing metabolic functions, raising body temperature, triggering the coagulation cascade, modifying hemodynamics and causing septic shock. Because of this toxicity, the removal of even minute amounts is essential for safe parenteral administration of drugs and also for septic shock patients' care. The absence of a general method for endotoxin removal from liquid interfaces urgently requires finding new methods and materials to overcome this gap. Nanostructured carbonized plant parts is a promising material that showed good adsorption properties due to its vast pore network and high surface area. The aim of this study was comparative measurement of endotoxin- and blood proteins-related adsorption rate and adsorption capacity for different carboneous materials produced at different temperatures and under different surface modifications. As a main surface modificator, positively cbarged polymer, polyethileneimine (PEl) was used. Activated carbon materials showed good adsorption properties for LPS and some proteins used in the experiments. During the batch experiments, several techniques (dust removal, autoclaving) were used and optimized for improving the material's adsorption behavior. Also, with the results obtained it was possible to differentiate the materials according to their adsorption capacity and kinetic characteristics. Modification of the surface apparently has not affected hemoglobin binding to the adsorbent's surface. Obtained adsorption isotherms can be used as a powerful tool for designing of future column-based setups for blood purification from LPS, which is especially important for septic shock treatment.
The sorption of LPS toxic shock by nanoparticles on base of carbonized vegetable raw materials
(2008)
Immobilization of lactobacillus on high temperature carbonizated vegetable raw material (rice husk, grape stones) increases their physiological activity and the quantity of the antibacterial metabolits, that consequently lead to increase of the antagonistic activity of lactobacillus. It is implies that the use of the nanosorbents for the attachment of the probiotical microorganisms are highly perspective for decision the important problems, such as the probiotical preparations delivery to the right address and their attachment to intestines mucosa with the following detoxication of gastro-intestinal tract and the normalization of it’s microecology. Besides that, thus, the received carbonizated nanoparticles have peculiar properties – ability to sorption of LPS toxical shock and, hence, to the detoxication of LPS.
Summary and Conclusions PCIs were clearly effective in terms of their antibacterial effects with the strains tested. This efficacy increased with the time the bacteries were exposed to PCIs. The bactericidal action has proved to be irreversible. PCIs were significantly less effective in shadowed areas. PCI exposure caused multiple protein damages as observed in SDS PAGE studies. There was no single but multiple molecular mechanism causing the bacterial death.
Recently, SHARP corporation has developed the world’s first "Plasma Cluster Ions® (PCI)" air purification technology, which uses plasma discharge to generate cluster ions. The new Plasma Cluster Device releases positive and negative ions into the air, which are harmless to humans and are able to decompose and deactivate airborne substances by chemical reactions. In the past, phenomenological tests on the efficacy of the PCI air purification technology on microbial cells have been conducted. In most cases, it has been shown that PCI demonstrated strongly pronounced killing effects on microorganisms. However, the particular mechanisms of PCI action still have to be uncovered.
Changes in intestinal microflora in rats induced by oral exposure to low lead (II) concentrations
(2015)
Recently, SHARP corporation has developed the world’s first “Plasma Cluster Ions (PCI)” air purification technology, which uses plasma discharge to generate cluster ions. The new plasma cluster device releases into the air positive and negative ions, which are harmless to humans and are able to decompose and deactivate airborne substances by chemical reactions. A lot of phenomenological tests of the PCI air purification technology on microbial cells have been conducted. And, in most cases, it has been shown that PCI demonstrate strongly pronounced killing effect. Although, the particular mechanisms of PCI action are still not evident. We studied variations in resistance to PCI among gram-positive airborne microorganisms, as well as some dose-dependent, spatial, cultural and biochemical properties of PCI action in respect of Staphylococcus spp, Enterococcus spp, Micrococcus spp.
On the model of musculocutaneous wound in rats, the effect of applicative sorption by carbonized rise shell (CRS) on the healing of festering wound was studied. It has been shown, that cytological changes end with rapid scar formation. The use of CRS at the period of severe purulent wound contributes to its favorable course, prevents the development of complications of the animals from sepsis.
Optical coherence tomography : a potential tool to predict premature rupture of fetal membranes
(2013)
Tests with palm tree leaves have just started yet and scan data are in the process to be analyzed. The final goal of future project for palm tree gender and species recognition will be to develop optical scanning technology to be applied to date palm tree leaves for in–situ screening purposes. Depending on the software used and the particular requirements of the users the technology potentially shall be able to identify palm tree diseases, palm tree gender, and species of young date palm trees by scanning leaves.
The demand of replacements for inoperable organs exceeds the amount of available organ transplants. Therefore, tissue engineering developed as a multidisciplinary field of research for autologous in-vitro organs. Such three dimensional tissue constructs request the application of a bioreactor. The UREPLACE bioreactor is used to grow cells on tubular collagen scaffolds OPTIMAIX Sponge 1 with a maximal length of 7 cm, in order to culture in vitro an adequate ureter replacement. With a rotating unit, (urothelial) cells can be placed homogeneously on the inner scaffold surface. Furthermore, a stimulation is combined with this bioreactor resulting in an orientation of muscle cells. These culturing methods request a precise control of several parameters and actuators. A combination of a LabBox and the suitable software LabVision is used to set and conduct parameters like rotation angles, velocities, pressures and other important cell culture values. The bioreactor was tested waterproof successfully. Furthermore, the temperature controlling was adjusted to 37 °C and the CO2 - concentration regulated to 5 %. Additionally, the pH step responses of several substances showed a perfect functioning of the designed flow chamber. All used software was tested and remained stable for several days.
Reconstructive surgery and tissue replacements like ureters or bladders reconstruction have been recently studied, taking into account growth and remodelling of cells since living cells are capable of growing, adapting, remodelling or degrading and restoring in order to deform and respond to stimuli. Hence, shapes of ureters or bladders and their microstructure change during growth and these changes strongly depend on external stimuli such as training. We present the mechanical stimulation of smooth muscle cells in a tubular fibrin-PVDFA scaffold and the modelling of the growth of tissue by stimuli. To this end, mechanotransduction was performed with a kyphoplasty balloon catheter that was guided through the lumen of the tubular structure. The bursting pressure was examined to compare the stability of the incubated tissue constructs. The results showed the significant changes on tissues with training by increasing the burst pressure as a characteristic mechanical property and the smooth muscle cells were more oriented with uniformly higher density. Besides, the computational growth models also exhibited the accurate tendencies of growth of the cells under different external stimuli. Such models may lead to design standards for the better layered tissue structure in reconstructing of tubular organs characterized as composite materials such as intestines, ureters and arteries.
Advances in polymer science have significantly increased polymer applications in life sciences. We report the use of free-standing, ultra-thin polydimethylsiloxane (PDMS) membranes, called CellDrum, as cell culture substrates for an in vitro wound model. Dermal fibroblast monolayers from 28- and 88-year-old donors were cultured on CellDrums. By using stainless steel balls, circular cell-free areas were created in the cell layer (wounding). Sinusoidal strain of 1 Hz, 5% strain, was applied to membranes for 30 min in 4 sessions. The gap circumference and closure rate of un-stretched samples (controls) and stretched samples were monitored over 4 days to investigate the effects of donor age and mechanical strain on wound closure. A significant decrease in gap circumference and an increase in gap closure rate were observed in trained samples from younger donors and control samples from older donors. In contrast, a significant decrease in gap closure rate and an increase in wound circumference were observed in the trained samples from older donors. Through these results, we propose the model of a cell monolayer on stretchable CellDrums as a practical tool for wound healing research. The combination of biomechanical cell loading in conjunction with analyses such as gene/protein expression seems promising beyond the scope published here.
Hypertension describes the pathological increase of blood pressure, which is most commonly associated with the increase of vascular wall stiffness [1]. Referring to the “Deutsche Bluthochdruck Liga” this pathology shows a growing trend in our aging society. In order to find novel pharmacological and probably personalized treatments, we want to present a functional approach to study biomechanical properties of a human aortic vascular model.
In this method review we will give an overview of recent studies which were carried out with the CellDrum technology [2] and underline the added value to already existing standard procedures known from the field of physiology.
Herein described CellDrum technology is a system to measure functional mechanical properties of cell monolayers and thin tissue constructs in-vitro. Additionally, the CellDrum enables to elucidate the mechanical response of cells to pharmacological drugs, toxins and vasoactive agents. Due to its highly flexible polymer support, cells can also be mechanically stimulated by steady and cyclic biaxial stretching.
Background/Aims: Common systems for the quantification of cellular contraction rely on animal-based models, complex experimental setups or indirect approaches. The herein presented CellDrum technology for testing mechanical tension of cellular monolayers and thin tissue constructs has the potential to scale-up mechanical testing towards medium-throughput analyses. Using hiPS-Cardiac Myocytes (hiPS-CMs) it represents a new perspective of drug testing and brings us closer to personalized drug medication. Methods: In the present study, monolayers of self-beating hiPS-CMs were grown on ultra-thin circular silicone membranes and deflect under the weight of the culture medium. Rhythmic contractions of the hiPS-CMs induced variations of the membrane deflection. The recorded contraction-relaxation-cycles were analyzed with respect to their amplitudes, durations, time integrals and frequencies. Besides unstimulated force and tensile stress, we investigated the effects of agonists and antagonists acting on Ca²⁺ channels (S-Bay K8644/verapamil) and Na⁺ channels (veratridine/lidocaine). Results: The measured data and simulations for pharmacologically unstimulated contraction resembled findings in native human heart tissue, while the pharmacological dose-response curves were highly accurate and consistent with reference data. Conclusion: We conclude that the combination of the CellDrum with hiPS-CMs offers a fast, facile and precise system for pharmacological, toxicological studies and offers new preclinical basic research potential.