Search results for: regenerative endodontic
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 172

Search results for: regenerative endodontic

142 Design of an Electric Vehicle Model with a Dynamo Drive Setup Using Model-Based Development (MBD) (EV Using MBD)

Authors: Gondu Vykunta Rao, Madhuri Bayya, Aruna Bharathi M., Paramesw Chidamparam, B. Murali

Abstract:

The increase in software content in today’s electric vehicles is increasing attention to having vast, unique topographies from low emission to high efficiency, whereas the chemical batteries have huge short comes, such as limited cycle life, power density, and cost. As for understanding and visualization, the companies are turning toward the virtual vehicle to test their design in software which is known as a simulation in the loop (SIL). In this project, in addition to the electric vehicle (EV) technology, we are adding a dynamo with the vehicle for regenerative braking. Traditionally the principle of dynamos is used in lighting the purpose of the bicycle. Here by using the same mechanism, we are running the vehicle as well as charging the vehicle from system-level simulation to the model in the loop and then to the Hardware in Loop (HIL) by using model-based development.

Keywords: electric vehicle, simulation in the loop (SIL), model in loop (MIL), hardware in loop (HIL), dynamos, model-based development (MBD), permanent magnet synchronous motor (PMSM), current control (CC), field-oriented control (FOC), regenerative braking

Procedia PDF Downloads 122
141 Cochlear Implants and the Emerging Therapies for Managing Hearing Loss

Authors: Hesham Kozou

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Sensorineural hearing loss (SNHL) poses a significant challenge due to limited access to the inner ear for therapies. Emerging treatments such as regenerative, genetic, and pharmacotherapies offer hope for addressing this condition. This study aims to highlight the potential of cochlear implants and emerging therapies in managing sensorineural hearing loss by improving access to the inner ear. The study is conducted through a review of relevant literature and research articles in the field of cochlear implants and emerging therapies for hearing loss. It outlines how advancements in cochlear implant technologies, electrodes, and surgical techniques can facilitate the delivery of therapies to the inner ear, potentially revolutionizing the treatment of sensorineural hearing loss. The study underscores the potential of cochlear implants and emerging therapies in revolutionizing the treatment landscape for sensorineural hearing loss, emphasizing the feasibility of curing this condition by leveraging technological advancements.

Keywords: therapies for hearing loss management, future of CI as a cochlear delivery channel, regenerative, genetic and pharmacotherapeutic management of hearing loss

Procedia PDF Downloads 48
140 Evaluation of Biological Seed Coating Technology On-Field Performance of Wheat in Regenerative Agriculture and Conventional Systems

Authors: S. Brain, P. J. Storer, H. Strydom, Z. M. Solaiman

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Increasing farmer awareness of soil health, the impact of agricultural management practices, and the requirement for high-quality agricultural produce are major factors driving the rapid adoption of biological seed treatments - currently valued globally at USD 1.5 billion. Biological seed coatings with multistrain plant beneficial microbial technology have the capability to affect plant establishment, growth, and development positively. These beneficial plant microbes can potentially increase soil health, plant yield, and nutrition – acting as bio fertilisers, rhizoremediators, phytostimulators, and stress modulators, and can ultimately reduce the overall use of agrichemicals. A field trial was conducted on MACE wheat in the central wheat belt of Western Australia to evaluate a proprietary seed coating technology (Langleys Bio-EnergeticTM Microbe blend (BMB)) on a conventional program (+/- BMB microbes) and a Regenerative Biomineral fertiliser program (+/- BMB microbes). The Conventional (+BMB) and Biomineral (+BMB) treated plants had no fungicide treatments and had no disease issues. Control (No fertiliser, No microbes), Conventional (No Microbes), and Biomineral (No Microbes) were treated with fungicides (seed dressing and foliar). From the research findings, compared to control and no microbe treatments, both the Conventional (+ BMB) and Biomineral (+ BMB) showed significant increases in Soil Carbon (SOC), Seed germination, nutrient use efficiency (NUE) of nitrogen, phosphate and mineral nutrients, grain mineral nutrient uptake, protein %, hectolitre weight, and fewer screenings, yield, and gross margins.

Keywords: biological seed coating, biomineral fertiliser, plant nutrition, regenerative and conventional agriculture

Procedia PDF Downloads 79
139 Numerical Analysis of Various V- rib Cross-section to Optimize Thermal Performance of the Rocket Engine

Authors: Hisham Elmouazen, Xiaobing Zhang

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In regenerative-cooled rocket engines, understanding the coolant behaviour within cooling channels is essential to enhance engine performance and maintain chamber walls at low temperatures. However, modelling and testing the rocket engine's cooling channels is challenging due to the high temperature of the chamber walls, supercritical flow, and high Reynolds number. Therefore, a numerical analysis of five different V-rib cross-sections to optimize rocket engine cooling channels' performance is developed and validated in this work. Three-dimensional CFD simulations are employed by the Shear Stress Transport (k- ω) turbulent model at Reynolds number 42,500. The study findings illustrate that the V-ribbed channel performance is optimized by 59.5% relative to the plain/flat channel. Additionally, the chamber wall temperature is decreased to 726.4 K, and the right-angle trapezoidal V-rib (Case 4) improves thermal augmentation up to 74.3 % with a slightly high friction factor.

Keywords: computational fluid dynamics CFD, regenerative-cooled system, thermal performance, V-rib cross-sections

Procedia PDF Downloads 75
138 Comparative Evaluation of a Dynamic Navigation System Versus a Three-Dimensional Microscope in Retrieving Separated Endodontic Files: An in Vitro Study

Authors: Mohammed H. Karim, Bestoon M. Faraj

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Introduction: instrument separation is a common challenge in the endodontic field. Various techniques and technologies have been developed to improve the retrieval success rate. This study aimed to compare the effectiveness of a Dynamic Navigation System (DNS) and a three-dimensional microscope in retrieving broken rotary NiTi files when using trepan burs and the extractor system. Materials and Methods: Thirty maxillary first bicuspids with sixty separate roots were split into two comparable groups based on a comprehensive Cone-Beam Computed Tomography (CBCT) analysis of the root length and curvature. After standardised access opening, glide paths, and patency attainment with the K file (sizes 10 and 15), the teeth were arranged on 3D models (three per quadrant, six per model). Subsequently, controlled-memory heat-treated NiTi rotary files (#25/0.04) were notched 4 mm from the tips and fractured at the apical third of the roots. The C-FR1 Endo file removal system was employed under both guidance to retrieve the fragments, and the success rate, canal aberration, treatment time and volumetric changes were measured. The statistical analysis was performed using IBM SPSS software at a significance level of 0.05. Results: The microscope-guided group had a higher success rate than the DNS guidance, but the difference was insignificant (p > 0.05). In addition, the microscope-guided drills resulted in a substantially lower proportion of canal aberration, required less time to retrieve the fragments and caused a minor change in the root canal volume (p < 0.05). Conclusion: Although dynamically guided trephining with the extractor can retrieve separated instruments, it is inferior to three-dimensional microscope guidance regarding treatment time, procedural errors, and volume change.

Keywords: dynamic navigation system, separated instruments retrieval, trephine burs and extractor system, three-dimensional video microscope

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137 Comparative Study of Sub-Critical and Supercritical ORC Applications for Exhaust Waste Heat Recovery

Authors: Buket Boz, Alvaro Diez

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Waste heat recovery by means of Organic Rankine Cycle is a promising technology for the recovery of engine exhaust heat. However, it is complex to find out the optimum cycle conditions with appropriate working fluids to match exhaust gas waste heat due to its high temperature. Hence, this paper focuses on comparing sub-critical and supercritical ORC conditions with eight working fluids on a combined diesel engine-ORC system. The model employs two ORC designs, Regenerative-ORC and Pre-Heating-Regenerative-ORC respectively. The thermodynamic calculations rely on the first and second law of thermodynamics, thermal efficiency and exergy destruction factors are the fundamental parameters evaluated. Additionally, in this study, environmental and safety, GWP (Global Warming Potential) and ODP (Ozone Depletion Potential), characteristic of the refrigerants are taken into consideration as evaluation criteria to define the optimal ORC configuration and conditions. Consequently, the studys outcomes reveal that supercritical ORCs with alkane and siloxane are more suitable for high temperature exhaust waste heat recovery in contrast to sub-critical conditions.

Keywords: internal combustion engine, organic Rankine cycle, waste heat recovery, working fluids

Procedia PDF Downloads 204
136 Performance Analysis of Organic Rankine Cycle Technology to Exploit Low-Grade Waste Heat to Power Generation in Indian Industry

Authors: Bipul Krishna Saha, Basab Chakraborty, Ashish Alex Sam, Parthasarathi Ghosh

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The demand for energy is cumulatively increasing with time.  Since the availability of conventional energy resources is dying out gradually, significant interest is being laid on searching for alternate energy resources and minimizing the wastage of energy in various fields.  In such perspective, low-grade waste heat from several industrial sources can be reused to generate electricity. The present work is to further the adoption of the Organic Rankine Cycle (ORC) technology in Indian industrial sector.  The present paper focuses on extending the previously reported idea to the next level through a comparative review with three different working fluids using practical data from an Indian industrial plant. For comprehensive study in the simulation platform of Aspen Hysys®, v8.6, the waste heat data has been collected from a current coke oven gas plant in India.  A parametric analysis of non-regenerative ORC and regenerative ORC is executed using the working fluids R-123, R-11 and R-21 for subcritical ORC system.  The primary goal is to determine the optimal working fluid considering various system parameters like turbine work output, obtained system efficiency, irreversibility rate and second law efficiency under applied multiple heat source temperature (160 °C- 180 °C).  Selection of the turbo-expanders is one of the most crucial tasks for low-temperature applications in ORC system. The present work is an attempt to make suitable recommendation for the appropriate configuration of the turbine. In a nutshell, this study justifies the proficiency of integrating the ORC technology in Indian perspective and also finds the appropriate parameter of all components integrated in ORC system for building up an ORC prototype.

Keywords: organic Rankine cycle, regenerative organic Rankine cycle, waste heat recovery, Indian industry

Procedia PDF Downloads 374
135 Micromechanical Compatibility Between Cells and Scaffold Mediates the Efficacy of Regenerative Medicine

Authors: Li Yang, Yang Song, Martin Y. M. Chiang

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Objective: To experimentally substantiate the micromechanical compatibility between cell and scaffold, in the regenerative medicine approach for restoring bone volume, is essential for phenotypic transitions Methods: Through nanotechnology and electrospinning process, nanofibrous scaffolds were fabricated to host dental follicle stem cells (DFSCs). Blends (50:50) of polycaprolactone (PCL) and silk fibroin (SF), mixed with various content of cellulose nanocrystals (CNC, up to 5% in weight), were electrospun to prepare nanofibrous scaffolds with heterogeneous microstructure in terms of fiber size. Colloidal probe atomic force microscopy (AFM) and conventional uniaxial tensile tests measured the scaffold stiffness at the micro-and macro-scale, respectively. The cell elastic modulus and cell-scaffold adhesive interaction (i.e., a chemical function) were examined through single-cell force spectroscopy using AFM. The quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to determine if the mechanotransduction signal (i.e., Yap1, Wwr2, Rac1, MAPK8, Ptk2 and Wnt5a) is upregulated by the scaffold stiffness at the micro-scale (cellular scale). Results: The presence of CNC produces fibrous scaffolds with a bimodal distribution of fiber diameter. This structural heterogeneity, which is CNC-composition dependent, remarkably modulates the mechanical functionality of scaffolds at microscale and macroscale simultaneously, but not the chemical functionality (i.e., only a single material property is varied). In in vitro tests, the osteogenic differentiation and gene expression associated with mechano-sensitive cell markers correlate to the degree of micromechanical compatibility between DFSCs and the scaffold. Conclusion: Cells require compliant scaffolds to encourage energetically favorable interactions for mechanotransduction, which are converted into changes in cellular biochemistry to direct the phenotypic evolution. The micromechanical compatibility is indeed important to the efficacy of regenerative medicine.

Keywords: phenotype transition, scaffold stiffness, electrospinning, cellulose nanocrystals, single-cell force spectroscopy

Procedia PDF Downloads 190
134 Thermal Performance of Reheat, Regenerative, Inter-Cooled Gas Turbine Cycle

Authors: Milind S. Patil, Purushottam S. Desale, Eknath R. Deore

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Thermal analysis of reheat, regenerative, inter-cooled gas turbine cycle is presented. Specific work output, thermal efficiency and SFC is simulated with respect to operating conditions. Analytical formulas were developed taking into account the effect of operational parameters like ambient temperature, compression ratio, compressor efficiency, turbine efficiency, regenerator effectiveness, pressure loss in inter cooling, reheating and regenerator. Calculations were made for wide range of parameters using engineering equation solver and the results were presented here. For pressure ratio of 12, regenerator effectiveness 0.95, and maximum turbine inlet temperature 1200 K, thermal efficiency decreases by 27% with increase in ambient temperature (278 K to 328 K). With decrease in regenerator effectiveness thermal efficiency decreases linearly. With increase in ambient temperature (278 K to 328 K) for the same maximum temperature and regenerator effectiveness SFC decreases up to a pressure ratio of 10 and then increases. Sharp rise in SFC is noted for higher ambient temperature. With increase in isentropic efficiency of compressor and turbine, thermal efficiency increases by about 40% for low ambient temperature (278 K to 298 K) however, for higher ambient temperature (308 K to 328 K) thermal efficiency increases by about 70%.

Keywords: gas turbine, reheating, regeneration, inter-cooled, thermal analysis

Procedia PDF Downloads 337
133 Biomimetic to Architectural Design for Increased Sustainability

Authors: Hamid Yazdani, Fatemeh Abbasi

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Biomimicry, where flora, fauna or entire ecosystems are emulated as a basis for design, is a growing area of research in the fields of architecture and engineering. This is due to both the fact that it is an inspirational source of possible new innovation and because of the potential it offers as a way to create a more sustainable and even regenerative built environment. The widespread and practical application of biomimicry as a design method remains however largely unrealised. A growing body of international research identifies various obstacles to the employment of biomimicry as an architectural design method. One barrier of particular note is the lack of a clear definition of the various approaches to biomimicry that designers can initially employ. Through a comparative literature review, and an examination of existing biomimetic technologies, this paper elaborates on distinct approaches to biomimetic design that have evolved. A framework for understanding the various forms of biomimicry has been developed, and is used to discuss the distinct advantages and disadvantages inherent in each as a design methodology. It is shown that these varied approaches may lead to different outcomes in terms of overall sustainability or regenerative potential. It is posited that a biomimetic approach to architectural design that incorporates an understanding of ecosystems could become a vehicle for creating a built environment that goes beyond simply sustaining current conditions to a restorative practice where the built environment becomes a vital component in the integration with and regeneration of natural ecosystems.

Keywords: biomimicry, bio-inspired design, ecology, ecomimicry, industrial ecology

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132 Developing Alternative Recovery Technology of Waste Heat in Automobile Factory

Authors: Kun-Ping Cheng, Dong-Shang Chang, Rou-Wen Wang

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Pre-treatment of automobile paint-shop procedures are the preparation of warm water rinsing tank, hot water rinsing tank, degreasing tank, phosphate tank. The conventional boiler steam fuel is natural gas, producing steam to supply the heat exchange of each tank sink. In this study, the high-frequency soldering economizer is developed for recovering waste heat in the automotive paint-shop (RTO, Regenerative Thermal Oxidation). The heat recovery rate of the new economizer is 20% to 30% higher than the conventional embedded heat pipe. The adaptive control system responded to both RTO furnace exhaust gas and heat demands. In order to maintain the temperature range of the tanks, pre-treatment tanks are directly heated by waste heat recovery device (gas-to-water heat exchanger) through the hot water cycle of heat transfer. The performance of developed waste heat recovery system shows the annual recovery achieved to 1,226,411,483 Kcal of heat (137.8 thousand cubic meters of natural gas). Boiler can reduce fuel consumption by 20 to 30 percent compared to without waste heat recovery. In order to alleviate environmental impacts, the temperature at the end of the flue is further reduced from 160 to 110°C. The innovative waste heat recovery is helpful to energy savings and sustainable environment.

Keywords: waste heat recovery system, sustainability, RTO (Regenerative Thermal Oxidation), economizer, automotive industry

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131 Comparative Study of Non-Identical Firearms with Priority to Repair Subject to Inspection

Authors: A. S. Grewal, R. S. Sangwan, Dharambir, Vikas Dhanda

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The purpose of this paper is to develop and analyze two reliability models for a system of non-identical firearms – one is standard firearm (called as original unit) and the other is a country-made firearm (called as duplicate /substandard unit). There is a single server who comes immediately to do inspection and repair whenever needed. On the failure of standard firearm, the server inspects the operative country-made firearm to see whether the unit is capable of performing the desired function well or not. If country-made firearm is not capable to do so, the operation of the system is stopped and server starts repair of the standard firearms immediately. However, no inspection is done at the failure of the country-made firearm as the country-made firearm alone is capable of performing the given task well. In model I, priority to repair the standard firearm is given in case system fails completely and country-made firearm is already under repair, whereas in model II there is no such priority. The failure and repair times of each unit are assumed to be independent and uncorrelated random variables. The distributions of failure time of the units are taken as negative exponential while that of repair and inspection times are general. By using semi-Markov process and regenerative point technique some econo-reliability measures are obtained. Graphs are plotted to compare the MTSF (mean time to system failure), availability and profit of the models for a particular case.

Keywords: non-identical firearms, inspection, priority to repair, semi-Markov process, regenerative point

Procedia PDF Downloads 426
130 Periodontal Disease or Cement Disease: New Frontier in the Treatment of Periodontal Disease in Dogs

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, cement disease, regenerative therapy

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129 Development of Chitosan/Dextran Gelatin Methacrylate Core/Shell 3D Scaffolds and Protein/Polycaprolactone Melt Electrowriting Meshes for Tissue Regeneration Applications

Authors: J. D. Cabral, E. Murray, P. Turner, E. Hewitt, A. Ali, M. McConnell

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Worldwide demand for organ replacement and tissue regeneration is progressively increasing. Three-dimensional (3D) bioprinting, where a physical construct is produced using computer-aided design, is a promising tool to advance the tissue engineering and regenerative medicine fields. In this paper we describe two different approaches to developing 3D bioprinted constructs for use in tissue regeneration. Bioink development is critical in achieving the 3D biofabrication of functional, regenerative tissues. Hydrogels, cross-linked macromolecules that absorb large amounts of water, have received widespread interest as bioinks due to their relevant soft tissue mechanics, biocompatibility, and tunability. In turn, not only is bioink optimisation crucial, but the creation of vascularized tissues remains a key challenge for the successful fabrication of thicker, more clinically relevant bioengineered tissues. Among the various methodologies, cell-laden hydrogels are regarded as a favorable approach; and when combined with novel core/shell 3D bioprinting technology, an innovative strategy towards creating new vessel-like structures. In this work, we investigate this cell-based approach by using human umbilical endothelial cells (HUVECs) entrapped in a viscoelastic chitosan/dextran (CD)-based core hydrogel, printed simulataneously along with a gelatin methacrylate (GelMA) shell. We have expanded beyond our previously reported FDA approved, commercialised, post-surgical CD hydrogel, Chitogel®, by functionalizing it with cell adhesion and proteolytic peptides in order to promote bone marrow-derived mesenchymal stem cell (immortalized BMSC cell line, hTERT) and HUVECs growth. The biocompatibility and biodegradability of these cell lines in a 3D bioprinted construct is demonstrated. Our studies show that particular peptide combinations crosslinked within the CD hydrogel was found to increase in vitro growth of BMSCs and HUVECs by more than two-fold. These gels were then used as a core bioink combined with the more mechanically robust, UV irradiated GelMA shell bioink, to create 3D regenerative, vessel-like scaffolds with high print fidelity. As well, microporous MEW scaffolds made from milk proteins blended with PCL were found to show promising bioactivity, exhibiting a significant increase in keratinocyte (HaCaTs) and fibroblast (normal human dermal fibroblasts, NhDFs) cell migration and proliferation when compared to PCL only scaffolds. In conclusion, our studies indicate that a peptide functionalized CD hydrogel bioink reinforced with a GelMA shell is biocompatible, biodegradable, and an appropriate cell delivery vehicle in the creation of regenerative 3D constructs. In addition, a novel 3D printing technique, melt electrowriting (MEW), which allows fabrication of micrometer fibre meshes, was used to 3D print polycaprolactone (PCL) and bioactive milk protein, lactorferrin (LF) and whey protein (WP), blended scaffolds for potential skin regeneration applications. MEW milk protein/PCL scaffolds exhibited high porosity characteristics, low overall biodegradation, and rapid protein release. Human fibroblasts and keratinocyte cells were seeded on to the scaffolds. Scaffolds containing high concentrations of LF and combined proteins (LF+WP) showed improved cell viability over time as compared to PCL only scaffolds. This research highlights two scaffolds made using two different 3D printing techniques using a combination of both natural and synthetic biomaterial components in order to create regenerative constructs as potential chronic wound treatments.

Keywords: biomaterials, hydrogels, regenerative medicine, 3D bioprinting

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128 Effect of Dual Wavelength Light Exposure on Regeneration of Dugesia dorotocephala

Authors: Zayedali Shaikh

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Increasingly now more than ever, UV damage brings with it a litany of minor deformities that can range from mild lesions and discoloring to cataracts and blindness. Pluripotent stem cells in planaria and human skin can be used to treat wounds and skin damage, with the primary limitations being inadequate growth factors. Photobiomodulation therapy in the form of low-intensity red light therapy has been proven to provide helpful benefits in the healing of skin that displays some of the symptoms of UV damage, such as burns and lesions, along with stimulating the proliferation of stem cells in recellularizing tissue. This paper puts forth an alternate means by which to treat the effects of UV damage using the freshwater planarian model system, Dugesia dorotocephala, known for its regenerative abilities and abundance of pluripotent stem cells, which allow for the rapid growth and repair of missing or damaged structures. Our work consisted of exposing planaria to different types of light: red light, blue light, white light, darkness, red and blue light together, UV light, and finally, red and UV light together. The primary focus of this research was on the red and UV lights, with six controls acting as metrics to compare our findings. Through computer-assisted morphological analysis, the results show that there is no significant difference in the rates of regeneration of planaria treated with simultaneous exposure to red and UV light versus planaria in darkness (p > .05), a representation of their preferred natural habitat. Our research suggests the viability of red-light therapy in actively combating UV damage and expediting the growth of epidermal stem cells by acting as another growth factor.

Keywords: regenerative medicine, stem cells, planaria, photobiomodulation

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127 Removal Capacity of Activated Carbon (AC) by Combining AC and Titanium Dioxide (TIO₂) in a Photocatalytically Regenerative Activated Carbon

Authors: Hanane Belayachi, Sarra Bourahla, Amel Belayachi, Fadela Nemchi, Mostefa Belhakem

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The most used techniques to remove pollutants from wastewater are adsorption onto activated carbon (AC) and oxidation using a photocatalyst slurry. The aim of this work is to eliminate such drawbacks by combining AC and titanium dioxide (TiO₂) in a photocatalytically Regenerative Activated Carbon. Anatase titania was deposited on powder-activated carbon made from grape seeds by the impregnation method, and then the composite photocatalyst was employed for the removal of reactive black 5, which is an anionic azo dye, from water. The AGS/TiO₂ was characterized by BET, MEB, RDX and optical absorption spectroscopy. The BET surface area and the pore structure of composite photocatalysts (AGS/TiO₂) and activated grape seeds (AGS) were evaluated from nitrogen adsorption data at 77 K in relation to process conditions. Our results indicate that the photocatalytic activity of AGS/TiO₂ was much higher than single-phase titania. The adsorption equilibrium of reactive black 5 from aqueous solutions on the examined materials was investigated. Langmuir, Freundlich, and Redlich–Petersen models were fitted to experimental equilibrium data, and their goodness of fit is compared. The degradation kinetics fitted well to the Langmuir-Hinselwood pseudo first order rate low. The photocatalytic activity of AGS/TiO₂ was much higher than virgin TiO₂. Chemical oxygen demand (COD) removal was measured at regular intervals to quantify the mineralization of the dye. Above 96% mineralization was observed. These results suggest that UV-irradiated TiO₂ immobilized on activated carbon may be considered an adequate process for the treatment of diluted colored textile wastewater.

Keywords: activated carbon, pollutant, catalysis, TiO₂

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126 Stromal Vascular Fraction Regenerative Potential in a Muscle Ischemia/Reperfusion Injury Mouse Model

Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

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Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

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125 Determining Cellular Biomarkers Sensitive to Low Damaging Exposure

Authors: Svetlana Guryeva, Inna Kornienko, Elena Petersen

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At present, translational medicine is a rapidly developing branch of biomedicine. The main idea of translational medicine is a practical application of fundamental research. One of the possible applications for translational medicine is researching therapies that improve human age-related organism condition. To fill the gap between experiments and clinical practice, it is necessary to create the standardized system for the investigation of different effects on cellular aging models. In this study, primary human fibroblasts derived from patients of different ages were used as a cellular aging model. The senescence-associated β-galactosidase activity, lipofuscin, γ-H2AX, the reactive oxygen species level, and cell death markers (annexin V/propidium iodide) were used as biomarkers of the cell functional state. The effects of damaging exposures (oxidative stress and heat shock), potential positive factors (metformin and acetaminophen), and their combinations were investigated using the described biomarkers. Oxidative stress and heat shock caused the increase in the levels of all biomarkers, and only the cells from young patients partly coped with stress 3 days after the exposures. Metformin improved the state of pretreatment cells from young and old patients. The acetaminophen did not show significant changes in the biomarker levels compare to the action of metformin. This study proved the opportunity to develop a standardized screening system based on biomarkers of the cell functional state to identify potential positive or negative effects of some physical and chemical exposures. Moreover, such a system can be useful for the aims of regenerative medicine to determine the effect of cell pretreatment before transplantation.

Keywords: biomarkers, primary fibroblasts, regenerative medicine, senescence, test system, translational medicine

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124 Identification of Genes Regulating Differentiation and Stemness of Human Mesenchymal Stem Cells for Gene Therapy in Regenerative Medicine

Authors: Tong Ming Liu

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Human mesenchymal stem cells (MSCs) represent the most used stem cells for clinical application, which have been used in over 1000 clinical trials to treat over 30 diseases due to multilineage differentiation potential, secretome and immunosuppression. Gene therapies of MSCs hold great promise in the treatment of many diseases due to enhanced MSC-based clinical outcomes. To identify genes for gene therapy of MSCs, by comparing gene expression profile before and after MSC differentiation following by functional screening, we have identified ZNF145 that regulated MSC differentiation. Forced expression of ZNF145 resulted in enhanced in vitro chondrogenesis of MSCs as an upstream factor of SOX9 and improved osteochondral repair upon implant into osteochondral defects in rodents. By comparing gene expression profile during differentiation of iPSCs toward MSCs, we also identified gene HOX regulating MSC stemness, which was much downregulated in late-passaged MSCs. Knockdown of this gene greatly compromised MSC stemness including abolished proliferation, decreased CFU-F, promoted senescence and reduced expression of cell surface antigens linked to the MSC phenotype. In addition, multi-linage differentiation was also greatly impaired. Notably, HOX overexpression resulted in improved multi-lineage differentiation. In the mechanism, HOX expression significantly deceased in late passage of MSCs compared with early passage of MSCs, correlating with MSC important genes. ChIP-seq data shown that HOX binds to genes related to MSC self-renewal and differentiation. Most importantly, most HOX binding sites are lost in late passage of MSCs. HOX exerts its effects by directing binding Twist1, one important gene of MSCs. The identification of the genes regulating MSC differentiation and stemness will provide and promising strategies for gene therapy of MSCs in regenerative medicine.

Keywords: mesenchymal stem cell, novel transcription factor, stemness, gene therapy, cartilage repair, signaling pathway

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123 Electromagnetic Tuned Mass Damper Approach for Regenerative Suspension

Authors: S. Kopylov, C. Z. Bo

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This study is aimed at exploring the possibility of energy recovery through the suppression of vibrations. The article describes design of electromagnetic dynamic damper. The magnetic part of the device performs the function of a tuned mass damper, thereby providing both energy regeneration and damping properties to the protected mass. According to the theory of tuned mass damper, equations of mathematical models were obtained. Then, under given properties of current system, amplitude frequency response was investigated. Therefore, main ideas and methods for further research were defined.

Keywords: electromagnetic damper, oscillations with two degrees of freedom, regeneration systems, tuned mass damper

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122 A Combination of Mesenchymal Stem Cells and Low-Intensity Ultrasound for Knee Meniscus Regeneration: A Preliminary Study

Authors: Mohammad Nasb, Muhammad Rehan, Chen Hong

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Background Meniscus defects critically alter knee function and lead to degenerative changes. Regenerative medicine applications including stem cell transplantation have showed a promising efficacy in finding alternatives to overcome traditional treatment limitations. However, stem cell therapy remains limited due to the substantially reduced viability and inhibitory microenvironment. Since tissue growth and repair are under the control of biochemical and mechanical signals, several approaches have recently been investigated (e.g., low intensity pulsed ultrasound [LIPUS]) to promote the regeneration process. This study employed LIPUS to improve growth and osteogenic differentiation of mesenchymal stem cells derived from human embryonic stem cells to improve the regeneration of meniscus tissue. Methodology: The Mesenchymal stromal cells (MSCs) were transplanted into the epicenter of the injured meniscus in rabbits, which were randomized into two main groups: a treatment group (n=32 New Zealand rabbits) including 4 subgroups of 8 rabbits in each subgroup (LIPUS treatment, MSC treatment, LIPUS with MSC and control), and a second group (n=9) to track implanted cells and their progeny using green fluorescence protein (GFP). GFP consists of the MSC and LIPUS-MSC combination subgroups. Rabbits were then subjected to histological, immunohistochemistry, and MRI assessment. Results: The quantity of the newly regenerated tissue in the combination treatment group that had Ultrasound irradiation after mesenchymal stem cells were better at all end points. Likewise, Tissue quality scores were also greater in knees treated with both approaches compared with controls and single treatment at all end points, achieving significance at twelve and twenty-four weeks [p < 0.05], and [p = 0.008] at twelve weeks. Differentiation into type-I and II collagen-expressing cells were higher in the combination group at up to twenty-four weeks. Conclusions: the combination of mesenchymal stem cells and LIPUS showed greater adhering to the sites of meniscus injury, differentiate into cells resembling meniscal fibrochondrocytes, and improve both quality and quantity of meniscal regeneration.

Keywords: stem cells, regenerative medicine, osteoarthritis, knee

Procedia PDF Downloads 119
121 Comparison of β-Cell Regenerative Potentials of Selected Sri Lankan Medicinal Plant Extracts in Alloxan-Induced Diabetic Rats

Authors: A. P. Attanayake, K. A. P. W. Jayatilaka, L. K. B. Mudduwa, C. Pathirana

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Triggering of β-cell regeneration is a recognized therapeutic strategy for the treatment of type 1 diabetes mellitus. One such approach to foster restoration and regeneration of β-cells is from exogenous natural extracts. The aim of the present study was to investigate and compare the β-cell regenerative potentials of the extracts of Spondias pinnata (Linn. f.) Kurz, Coccinia grandis (L.) Voigt and Gmelina arborea Roxb. in alloxan induced diabetic rats. Wistar rats were divided in to six groups (n=6); healthy untreated rats, alloxan induced diabetic untreated rats (150 mg/kg, ip), diabetic rats receiving the extracts of S. pinnata (1.0 g/kg), C. grandis (0.75 g/kg), G. arobrea (1.00 g/kg) and diabetic rats receiving glibenclamide (0.5 mg/kg) for 30 days. The assessment of selected biochemical parameters, histopathology and immunohistochemistry in the pancreatic tissue were done on the 30th day. The reduction in the percentage of HbA1C was in the decreasing order of C. grandis (35%), G. arborea (31%) and S. pinnata (29%) in alloxan induced diabetic rats (p< 0.05). The concentration of serum fructosamine, insulin and C-peptide were decreased significantly in a decreasing order of C. grandis (30%, 72%, 51%), G. arborea (25%, 44%, 44%) and S. pinnata (27%, 34%, 24%) in alloxan induced diabetic rats (p < 0.05). The extent of β-cell regeneration was in the decreasing order of C. grandis, G. arborea, S. pinnata reflected through the increased percentage of insulin secreting β-cells in alloxan induced diabetic rats. The extract of C. grandis produced the highest degree of β-cell regeneration demonstrated through an increase in the number of islets and percentage of the insulin secreting β-cells (75%) in the pancreas of diabetic rats (p < 0.05). Further the C. grandis extract produced a significant increase in mean profile diameter in small (118%), average (10%), and large (13%) islets as compared with diabetic control rats respectively. However, statistically significant increase in the islet profile diameter was shown only in average (2%) and large (5%) islets in the G. arborea extract treated rats and large islets (5%) in S. pinnata extract treated diabetic rats (p < 0.05). The β-cell regeneration potency was in the decreasing order of C. grandis (0.75 g/kg), G. arborea (1.00 g/kg) and S. pinnata (1.00 g/kg) in alloxan induced diabetic rats. The three plant extracts may be useful as natural agents of triggering the β-cell regeneration in the management of type 1 diabetes mellitus.

Keywords: alloxan-induced diabetic rats, β-cell regeneration, histopathology, immunohistochemistry

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120 Push-Out Bond Strength of Two Root-End Filling Materials in Root-End Cavities Prepared by Er,Cr: YSGG Laser or Ultrasonic Technique

Authors: Noushin Shokouhinejad, Hasan Razmi, Reza Fekrazad, Saeed Asgary, Ammar Neshati, Hadi Assadian, Sanam Kheirieh

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This study compared the push-out bond strength of mineral trioxide aggregate (MTA) and a new endodontic cement (NEC) as root-end filling materials in root-end cavities prepared by ultrasonic technique (US) or Er,Cr:YSGG laser (L). Eighty single-rooted extracted human teeth were endodontically treated, apicectomised and randomly divided into four following groups (n = 20): US/MTA, US/NEC, L/MTA and L/NEC. In US/MTA and US/NEC groups, rooted cavities were prepared with ultrasonic retrotip and filled with MTA and NEC, respectively. In L/MTA and L/NEC groups, root-end cavities were prepared using Er, Cr:YSGG laser and filled with MTA and NEC, respectively. Each root was cut apically to create a 2 mm-thick root slice for measurement of bond strength using a universal testing machine. Then, all slices were examined to determine the mode of bond failure. Data were analysed using two-way ANOVA. Root-end filling materials showed significantly higher bond strength in root-end cavities prepared using the ultrasonic technique (US/MTA and US/NEC) (P < 0.001). The bond strengths of MTA and NEC did not differ significantly. The failure modes were mainly adhesive for MTA, but cohesive for NEC. In conclusion, bond strengths of MTA and NEC to root-end cavities were comparable and higher in ultrasonically prepared cavities.

Keywords: bond strength, Er, Cr:YSGG laser, MTA, NEC, root-end cavity

Procedia PDF Downloads 345
119 Personalized Tissues and Organs Replacement – a Peek into the Future

Authors: Asaf Toker

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Matricelf developed a technology that enables the production of autologous engineered tissue composed of matrix and cells derived from patients Omentum biopsy. The platform showed remarkable pre-clinical results for several medical conditions. The company recently licensed the technology that enabled scientist at Tel Aviv university that 3D printed a human heart from human cells and matrix for the first time in human history. The company plans to conduct its first human clinical trial for Acute Spinal Cord Injury (SCI) early in 2023.

Keywords: tissue engineering, regenerative medicine, spinal Cord Injury, autologous implants, iPSC

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118 The System for Root Canal Length Measurement Based on Multifrequency Impedance Method

Authors: Zheng Zhang, Xin Chen, Guoqing Ding

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Electronic apex locators (EAL) has been widely used clinically for measuring root canal working length with high accuracy, which is crucial for successful endodontic treatment. In order to maintain high accuracy in different measurement environments, this study presented a system for root canal length measurement based on multifrequency impedance method. This measuring system can generate a sweep current with frequencies from 100 Hz to 1 MHz through a direct digital synthesizer. Multiple impedance ratios with different combinations of frequencies were obtained and transmitted by an analog-to-digital converter and several of them with representatives will be selected after data process. The system analyzed the functional relationship between these impedance ratios and the distance between the file and the apex with statistics by measuring plenty of teeth. The position of the apical foramen can be determined by the statistical model using these impedance ratios. The experimental results revealed that the accuracy of the system based on multifrequency impedance ratios method to determine the position of the apical foramen was higher than the dual-frequency impedance ratio method. Besides that, for more complex measurement environments, the performance of the system was more stable.

Keywords: root canal length, apex locator, multifrequency impedance, sweep frequency

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117 Regenerative Agriculture Standing at the Intersection of Design, Mycology, and Soil Fertility

Authors: Andrew Gennett

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Designing for fungal development means embracing the symbiotic relationship between the living system and built environment. The potential of mycelium post-colonization is explored for the fabrication of advanced pure mycelium products, going beyond the conventional methods of aggregating materials. Fruiting induction imparts desired material properties such as enhanced environmental resistance. Production approach allows for simultaneous generation of multiple products while scaling up raw materials supply suitable for architectural applications. The following work explores the integration of fungal environmental perception with computational design of built fruiting chambers. Polyporales, are classified by their porous reproductive tissues supported by a wood-like context tissue covered by a hard waterproofing coat of hydrobpobins. Persisting for years in the wild, these species represent material properties that would be highly desired in moving beyond flat sheets of arial mycelium as with leather or bacon applications. Understanding the inherent environmental perception of fungi has become the basis for working with and inducing desired hyphal differentiation. Working within the native signal interpretation of a mycelium mass during fruiting induction provides the means to apply textures and color to the final finishing coat. A delicate interplay between meeting human-centered goals while designing around natural processes of living systems represents a blend of art and science. Architecturally, physical simulations inform model design for simple modular fruiting chambers that change as fungal growth progresses, while biological life science principles describe the internal computations occurring within the fungal hyphae. First, a form filling phase of growth is controlled by growth chamber environment. Second, an initiation phase of growth forms the final exterior finishing texture. Hyphal densification induces cellular cascades, in turn producing the classical hardened cuticle, UV protective molecule production, as well, as waterproofing finish. Upon fruiting process completion, the fully colonized spent substrate holds considerable value and is not considered waste. Instead, it becomes a valuable resource in the next cycle of production scale-up. However, the acquisition of new substrate resources poses a critical question, particularly as these resources become increasingly scarce. Pursuing a regenerative design paradigm from the environmental perspective, the usage of “agricultural waste” for architectural materials would prove a continuation of the destructive practices established by the previous industrial regime. For these residues from fields and forests serve a vital ecological role protecting the soil surface in combating erosion while reducing evaporation and fostering a biologically diverse food web. Instead, urban centers have been identified as abundant sources of new substrate material. Diverting the waste from secondary locations such as food processing centers, papers mills, and recycling facilities not only reduces landfill burden but leverages the latent value of these waste steams as precious resources for mycelium cultivation. In conclusion, working with living systems through innovative built environments for fungal development, provides the needed gain of function and resilience of mycelium products. The next generation of sustainable fungal products will go beyond the current binding process, with a focus upon reducing landfill burden from urban centers. In final considerations, biophilic material builds to an ecologically regenerative recycling production cycle.

Keywords: regenerative agriculture, mycelium fabrication, growth chamber design, sustainable resource acquisition, fungal morphogenesis, soil fertility

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116 Prospects of Regenerative Medicine with Human Allogeneic Adipose Tissue-Derived Mesenchymal Stem Cell Sheets: Achievements and Future Outlook in Clinical Trials for Myopic Chorioretinal Atrophy

Authors: Norimichi Nagano, Yoshio Hirano, Tsutomu Yasukawa

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Mesenchymal stem cells are thought to confer neuroprotection, facilitate tissue regeneration and exert their effects on retinal degenerative diseases, however, adverse events such as proliferative vitreoretinopathy and preretinal membrane disease associated with cell suspension transplantation have also been reported. We have recently developed human (allogeneic) adipose tissue-derived mesenchymal stem cell (adMSC) sheets through our proprietary sheet transformation technique, which could potentially mitigate these adverse events. To clarify the properties of our adMSC sheets named PAL-222, we performed in vitro studies such as viability testing, cytokine secretions by ELISA, immunohistochemical study, and migration assay. The viability of the cells exceeded 70%. Vascular Endothelial Growth Factor (VEGF) and Pigment Epithelium-Derived Factor (PEDF), which are quite important cytokines for the retinal area, were observed. PAL-222 expressed type I collagen, a strength marker, type IV collagen, a marker of the basement membrane, and elastin, an elasticity marker. Finally, the migration assay was performed and showed negative, which means that PAL-222 is stably kept in the topical area and does not come to pieces. Next, to evaluate the efficacy in vivo, we transplanted PAL-222 into the subretinal space of the eye of Royal College of Surgeons rats with congenital retinal degeneration and assessed it for three weeks after transplantation. We confirmed that PAL-222 suppressed the decrease in the thickness of the outer nuclear layer, which means that the photoreceptor protective effect treated with PAL-222 was significantly higher than that in the sham group. (p < 0.01). This finding demonstrates that PAL-222 showed their retinoprotective effect in a model of congenital retinal degeneration. As the study suggested the efficacy of PAL-222 in both in vitro and in vivo studies, we are presently engaged in clinical trials of PAL-222 for myopic chorioretinal atrophy, which is one of the retinal degenerative diseases, for the purpose of regenerative medicine.

Keywords: cell sheet, clinical trial, mesenchymal stem cell, myopic chorioretinal atrophy

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115 Nanowire Substrate to Control Differentiation of Mesenchymal Stem Cells

Authors: Ainur Sharip, Jose E. Perez, Nouf Alsharif, Aldo I. M. Bandeas, Enzo D. Fabrizio, Timothy Ravasi, Jasmeen S. Merzaban, Jürgen Kosel

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Bone marrow-derived human mesenchymal stem cells (MSCs) are attractive candidates for tissue engineering and regenerative medicine, due to their ability to differentiate into osteoblasts, chondrocytes or adipocytes. Differentiation is influenced by biochemical and biophysical stimuli provided by the microenvironment of the cell. Thus, altering the mechanical characteristics of a cell culture scaffold can directly influence a cell’s microenvironment and lead to stem cell differentiation. Mesenchymal stem cells were cultured on densely packed, vertically aligned magnetic iron nanowires (NWs) and the effect of NWs on the cell cytoskeleton rearrangement and differentiation were studied. An electrochemical deposition method was employed to fabricate NWs into nanoporous alumina templates, followed by a partial release to reveal the NW array. This created a cell growth substrate with free-standing NWs. The Fe NWs possessed a length of 2-3 µm, with each NW having a diameter of 33 nm on average. Mechanical stimuli generated by the physical movement of these iron NWs, in response to a magnetic field, can stimulate osteogenic differentiation. Induction of osteogenesis was estimated using an osteogenic marker, osteopontin, and a reduction of stem cell markers, CD73 and CD105. MSCs were grown on the NWs, and fluorescent microscopy was employed to monitor the expression of markers. A magnetic field with an intensity of 250 mT and a frequency of 0.1 Hz was applied for 12 hours/day over a period of one week and two weeks. The magnetically activated substrate enhanced the osteogenic differentiation of the MSCs compared to the culture conditions without magnetic field. Quantification of the osteopontin signal revealed approximately a seven-fold increase in the expression of this protein after two weeks of culture. Immunostaining staining against CD73 and CD105 revealed the expression of antibodies at the earlier time point (two days) and a considerable reduction after one-week exposure to a magnetic field. Overall, these results demonstrate the application of a magnetic NW substrate in stimulating the osteogenic differentiation of MSCs. This method significantly decreases the time needed to induce osteogenic differentiation compared to commercial biochemical methods, such as osteogenic differentiation kits, that usually require more than two weeks. Contact-free stimulation of MSC differentiation using a magnetic field has potential uses in tissue engineering, regenerative medicine, and bone formation therapies.

Keywords: cell substrate, magnetic nanowire, mesenchymal stem cell, stem cell differentiation

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114 Precursor Muscle Cell’s Phenotype under Compression in a Biomimetic Mechanical Niche

Authors: Fatemeh Abbasi, Arne Hofemeier, Timo Betz

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Muscle growth and regeneration critically depend on satellite cells (SCs) which are muscle stem cells located between the basal lamina and myofibres. Upon damage, SCs become activated, enter the cell cycle, and give rise to myoblasts that form new myofibres, while a sub-population self-renew and re-populate the muscle stem cell niche. In aged muscle as well as in certain muscle diseases such as muscular dystrophy, some of the SCs lose their regenerative ability. Although it is demonstrated that the chemical composition of SCs quiescent niche is different from the activated niche, the mechanism initially activated in the SCs remains unknown. While extensive research efforts focused on potential chemical activation, no such factor has been identified to the author’s best knowledge. However, it is substantiated that niche mechanics affects SCs behaviors, such as stemness and engraftment. We hypothesize that mechanical stress in the healthy niche (homeostasis) is different from the regenerative niche and that this difference could serve as an early signal activating SCs upon fiber damage. To investigate this hypothesis, we develop a biomimetic system to reconstitute both, the mechanical and the chemical environment of the SC niche. Cells will be confined between two elastic polyacrylamide (PAA) hydrogels with controlled elastic moduli and functionalized surface chemistry. By controlling the distance between the PAA hydrogel surfaces, we vary the compression forces exerted by the substrates on the cells, while the lateral displacement of the upper hydrogel will create controlled shear forces. To establish such a system, a simplified system is presented. We engineered a sandwich-like configuration of two elastic PAA layer with stiffnesses between 1 and 10 kPa and confined a precursor myoblast cell line (C2C12) in between these layers. Our initial observations in this sandwich model indicate that C2C12 cells show different behaviors under mechanical compression if compared to a control one-layer gel without compression. Interestingly, this behavior is stiffness-dependent. While the shape of C2C12 cells in the sandwich consisting of two stiff (10 kPa) layers was much more elongated, showing almost a neuronal phenotype, the cell shape in a sandwich situation consisting of one stiff and one soft (1 kPa) layer was more spherical. Surprisingly, even in proliferation medium and at very low cell density, the sandwich situation stimulated cell differentiation with increased striation and myofibre formation. Such behavior is commonly found for confluent cells in differentiation medium. These results suggest that mechanical changes in stiffness and applied pressure might be a relevant stimulation for changes in muscle cell behavior.

Keywords: C2C12 cells, compression, force, satellite cells, skeletal muscle

Procedia PDF Downloads 124
113 Single Cell Sorter Driven by Resonance Vibration of Cell Culture Substrate

Authors: Misa Nakao, Yuta Kurashina, Chikahiro Imashiro, Kenjiro Takemura

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The Research Goal: With the growing demand for regenerative medicine, an effective mass cell culture process is required. In a repetitive subculture process for proliferating cells, preparing single cell suspension which does not contain any cell aggregates is highly required because cell aggregates often raise various undesirable phenomena, e.g., apoptosis and decrease of cell proliferation. Since cell aggregates often occur in cell suspension during conventional subculture processes, this study proposes a single cell sorter driven by a resonance vibration of a cell culture substrate. The Method and the Result: The single cell sorter is simply composed of a cell culture substrate and a glass pipe vertically placed against the cell culture substrate with a certain gap corresponding to a cell diameter. The cell culture substrate is made of biocompatible stainless steel with a piezoelectric ceramic disk glued to the bottom side. Applying AC voltage to the piezoelectric ceramic disk, an out-of-plane resonance vibration with a single nodal circle of the cell culture substrate can be excited at 5.5 kHz. By doing so, acoustic radiation force is emitted, and then cell suspension containing only single cells is pumped into the pipe and collected. This single cell sorter is effective to collect single cells selectively in spite of its quite simple structure. We collected C2C12 myoblast cell suspension by the single cell sorter with the vibration amplitude of 12 µmp-p and evaluated the ratio of single cells in number against the entire cells in the suspension. Additionally, we cultured the collected cells for 72 hrs and measured the number of cells after the cultivation in order to evaluate their proliferation. As a control sample, we also collected cell suspension by conventional pipetting, and evaluated the ratio of single cells and the number of cells after the 72-hour cultivation. The ratio of single cells in the cell suspension collected by the single cell sorter was 98.2%. This ratio was 9.6% higher than that collected by conventional pipetting (statistically significant). Moreover, the number of cells cultured for 72 hrs after the collection by the single cell sorter yielded statistically more cells than that collected by pipetting, resulting in a 13.6% increase in proliferated cells. These results suggest that the cell suspension collected by the single cell sorter driven by the resonance vibration hardly contains cell aggregates whose diameter is larger than the gap between the cell culture substrate and the pipe. Consequently, the cell suspension collected by the single cell sorter maintains high cell proliferation. Conclusions: In this study, we developed a single cell sorter capable of sorting and pumping single cells by a resonance vibration of a cell culture substrate. The experimental results show the single cell sorter collects single cell suspension which hardly contains cell aggregates. Furthermore, the collected cells show higher proliferation than that of cells collected by conventional pipetting. This means the resonance vibration of the cell culture substrate can benefit us with the increase in efficiency of mass cell culture process for clinical applications.

Keywords: acoustic radiation force, cell proliferation, regenerative medicine, resonance vibration, single cell sorter

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