Search results for: gelatin
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 107

Search results for: gelatin

47 Effect of Various Capping Agents on Photocatalytic, Antibacterial and Antibiofilm of ZnO Nanoparticles

Authors: K. Akhil, J. Jayakumar, S. Sudheer Khan

Abstract:

Zinc oxide nanoparticles (ZnO NPs) are extensively used in a wide variety of commercial products including sunscreen, textile and paints. The present study evaluated the effect of surface capping agents including polyethylene glycol (EG), gelatin, polyvinyl alcohol(PVA) and poly vinyl pyrrolidone(PVP) on photocatalytic activity of ZnO NPs. The particles were also tested for its antibacterial and antibiofilm activity against Staphylococcus aureus (MTCC 3160) and Pseudomonas aeruginosa (MTCC 1688). Preliminary characterization was done by UV-Visible spectroscopy. Electron microscopic analysis showed that the particles were hexagonal in shape. The hydrodynamic size distribution was analyzed by using dynamic light scattering method and crystalline nature was determined by X-Ray diffraction method.

Keywords: antibacterial, antibiofilm, capping agents, photodegradation, surface coating, zinc oxide nanoparticles

Procedia PDF Downloads 233
46 Durable Phantom Production Identical to Breast Tissue for Use in Breast Cancer Detection Research Studies

Authors: Hayrettin Eroglu, Adem Kara

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Recently there has been significant attention given to imaging of the biological tissues via microwave imaging techniques. In this study, a phantom for the test and calibration of Microwave imaging used in detecting unhealthy breast structure or tumors was produced by using sol gel method. The liquid and gel phantoms being used nowadays are not durable due to evaporation and their organic ingredients, hence a new design was proposed. This phantom was fabricated from materials that were widely available (water, salt, gelatin, and glycerol) and was easy to make. This phantom was aimed to be better from the ones already proposed in the literature in terms of its durability and stability. S Parameters of phantom was measured with 1-18 GHz Probe Kit and permittivity was calculated via Debye method in “85070” commercial software. One, three, and five-week measurements were taken for this phantom. Finally, it was verified that measurement results were very close to the real biological tissue measurement results.

Keywords: phantom, breast tissue, cancer, microwave imaging

Procedia PDF Downloads 324
45 Encapsulation of Volatile Citronella Essential oil by Coacervation: Efficiency and Release Kinetic Study

Authors: Rafeqah Raslan, Mastura AbdManaf, Junaidah Jai, Istikamah Subuki, Ana Najwa Mustapa

Abstract:

The volatile citronella essential oil was encapsulated by simple coacervation and complex coacervation using gum Arabic and gelatin as wall material. Glutaraldehyde was used in the methodology as crosslinking agent. The citronella standard calibration graph was developed with R2 equal to 0.9523 for the accurate determination of encapsulation efficiency and release study. The release kinetic was analyzed based on Fick’s law of diffusion for polymeric system and linear graph of log fraction release over log time was constructed to determine the release rate constant, k and diffusion coefficient, n. Both coacervation methods in the present study produce encapsulation efficiency around 94%. The capsules morphology analysis supported the release kinetic mechanisms of produced capsules for both coacervation process.

Keywords: simple coacervation, complex coacervation, encapsulation efficiency, release kinetic study

Procedia PDF Downloads 287
44 Influence of Natural Gum on Curcumin Supersaturationin Gastrointestinal Fluids

Authors: Patcharawalai Jaisamut, Kamonthip Wiwattanawongsa, Ruedeekorn Wiwattanapatapee

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Supersaturation of drugs in the gastrointestinal tract is one approach to increase the absorption of poorly water-soluble drugs. The stabilization of a supersaturated state was achieved by adding precipitation inhibitors that may act through a variety of mechanisms.In this study, the effect of the natural gums, acacia, gelatin, pectin and tragacanth on curcumin supersaturation in simulated gastric fluid (SGF) (pH 1.2), fasted state simulated gastric fluid (FaSSGF) (pH 1.6), and simulated intestinal fluid (SIF) (pH 6.8)was investigated. The results indicated that all natural gums significantly increased the curcum insolubility (about 1.2-6-fold)when compared to the absence of gum, and assisted in maintaining the supersaturated drug solution. Among the tested gums, pectin at 3% w/w was the best precipitation inhibitor with a significant increase in the degree of supersaturation about 3-fold in SGF, 2.4-fold in FaSSGF and 2-fold in SIF.

Keywords: curcumin, solubility, supersaturation, precipitation inhibitor

Procedia PDF Downloads 322
43 Hydrodynamics of Wound Ballistics

Authors: Harpreet Kaur, Er. Arjun, Kirandeep Kaur, P. K. Mittal

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Simulation of a human body from a 20% gelatin & 80% water mixture is examined from a wound ballistics point of view. Parameters such as incapacitation energy & temporary to permanent cavity size & tools of hydrodynamics have been employed to arrive at a model of the human body similar to the one adopted by NATO. Calculations using equations of motion yield a value of 339 µs in which a temporary cavity with maximum size settles down to a permanent cavity. This occurs for 10mm size bullets & settles down to a permanent cavity in the case of 4 different bullets, i.e., 5.45, 5.56, 7.62,10 mm sizes. The obtained results are in excellent agreement with the body as a right circular cylinder of 15 cm height & 10 cm diameter. An effort is made here in this work to present a sound theoretical base to parameters commonly used in wound ballistics from field experience discussed by Col Coats & Major Beyer.

Keywords: gelatine, gunshot, hydrodynamic model, oscillation time, temporary and permanent cavity, wound ballistics

Procedia PDF Downloads 37
42 Synthesis of Nickel Oxide Nanoparticles in Presence of Sodium Dodecyl Sulphate

Authors: Fereshteh Chekin, Sepideh Sadeghi

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Nickel nanoparticles have attracted much attention because of applications in catalysis, medical diagnostics and magnetic applications. In this work, we reported a simple and low-cost procedure to synthesize nickel oxide nanoparticles (NiO-NPs) by using sodium dodecyl sulphate (SDS) and gelatin as stabilizer. The synthesized NiO-NPs were characterized by a variety of means such as transmission electron microscope (TEM), powder X-ray diffraction (XRD), scanning electron microscope (SEM) and UV-vis spectroscopy. The results show that the NiO nanoparticles with high crystalline can be obtained using this simple method. The grain size measured by TEM was 16 in presence of SDS, which agrees well with the XRD data. SDS plays an important role in the formation of the NiO nanoparticles. Moreover, the NiO nanoparticles have been used as a solid phase catalyst for the decomposition of hydrazine hydrate at room temperatures. The decomposition process has been monitored by UV–vis analysis. The present study showed that nanoparticles are not poisoned after their repeated use in decomposition of hydrazine.

Keywords: nickel oxide nanoparticles, sodium dodecyl sulphate, synthesis, stabilizer

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41 Anti-Angiogenic and Anti-Metastatic Effect of Aqueous Fraction from Euchelus Asper Methanolic Extract

Authors: Sweta Agrawal, Sachin Chaugule, Gargi Rane, Shashank More, Madhavi Indap

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Angiogenesis and metastasis are two of the most important hallmarks of cancer. Hence, most of the cancer therapies nowadays are multi-targeted so as to reduce resistance and have better efficacy. As synthetic molecules arise with a burden of their toxicities and side-effects, more and more research is being focussed on exploiting the vast natural resources of drugs, in the form of plants and animals. Although, the idea of using marine organisms as a source of pharmaceuticals is not new, the pace at which marine drugs are being discovered, has definitely up surged! In the present study, we have assessed the anti-angiogenic and in vitro anti-metastatic activity of aqueous fraction from the extract of marine gastropod Euchelus asper. The soft body of Euchelus Asper was extracted with methanol and named EAME. Partition chromatography of EAME gave three fractions EAME I, II and III. Biochemical analysis revealed the presence of proteins in EAME III. Preliminary analysis had revealed the anti-angiogenic activity was exhibited by EAME III out of the three fractions. Hereafter, EAME III (concentration 25µg/ml-400µg/ml) was tested on chick chorioallantoic membrane (CAM) model for the detailed analysis of its potential anti-angiogenic effect. In vitro testing of the fraction (concentration 0.25µg/ml - 1µg/ml), involved cytotoxicity by SRB assay, cell cycle analysis by flow cytometry and anti-proliferative effect by scratch wound healing assay on A549 lung carcinoma cells. Apart from this, a portion of treated CAM as well as conditioned medium from treated A549 were subjected to gelatin zymography for assessment of matrix metalloproteinases MMP-2 and MMP-9 levels. Our results revealed that EAME III exhibited significant anti-angiogenic activity on CAM which was also supported by histological observations. During histological studies of CAM, it was found that EAME III caused reduction in angiogenesis by altering the extracellular matrix of the CAM membrane. In vitro analysis disclosed that EAME III exhibited moderate cytotoxic effect on A549 cells and its effect was not dose-dependent. The results of flow cytometry confirmed that EAME III caused cell cycle arrest in A549 cell line as almost all of the treated cells were found in G1 phase. Further, the migration and proliferation of A549 was significantly reduced by EAME III as observed from the scratch wound assay. Moreover, Gelatin zymography analysis revealed that EAME III caused suppression of MMP-2 in CAM membrane and reduced MMP-9 and MMP-2 expression in A549 cells. This verified that the anti-angiogenic and anti-metastatic effects of EAME III were correlated with the suppression of MMP-2 and -9. To conclude, EAME III shows dual anti-tumour action by reducing angiogenesis and exerting anti-metastatic effect on lung cancer cells, thus it has the potential to be used as an anti-cancer agent against lung carcinoma.

Keywords: angiogenesis, anti-cancer, marine drugs, matrix metalloproteinases

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40 Cryotopic Macroporous Polymeric Matrices for Regenerative Medicine and Tissue Engineering Applications

Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

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Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

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39 Assessment of Highly Sensitive Dielectric Modulated GaN-FinFET for Label-Free Biosensing Applications

Authors: Ajay Kumar, Neha Gupta

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This work presents the sensitivity assessment of Gallium Nitride (GaN) material-based FinFET by dielectric modulation in the nanocavity gap for label-free biosensing applications. The significant deflection is observed in the electrical characteristics such as drain current (ID), transconductance (gm), surface potential, energy band profile, electric field, sub-threshold slope (SS), and threshold voltage (Vth) in the presence of biomolecules owing to GaN material. Further, the device sensitivity is evaluated to identify the effectiveness of the proposed biosensor and its capability to detect the biomolecules with high precision or accuracy. Higher sensitivity is observed for Gelatin (k=12) in terms of on-current (SION), threshold voltage (SVth), and switching ratio (SSR) by 104.88%, 82.12%, and 119.73%, respectively. This work is performed using a powerful tool 3D Sentaurus TCAD using a well-calibrated structure. All the results pave the way for GaN-FinFET as a viable candidate for label-free dielectric modulated biosensor applications.

Keywords: biosensor, biomolecules, FinFET, sensitivity

Procedia PDF Downloads 154
38 Biodegradable Cross-Linked Composite Hydrogels Enriched with Small Molecule for Osteochondral Regeneration

Authors: Elena I. Oprita, Oana Craciunescu, Rodica Tatia, Teodora Ciucan, Reka Barabas, Orsolya Raduly, Anca Oancea

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Healing of osteochondral defects requires repair of the damaged articular cartilage, the underlying subchondral bone and the interface between these tissues (the functional calcified layer). For this purpose, developing a single monophasic scaffold that can regenerate two specific lineages (cartilage and bone) becomes a challenge. The aim of this work was to develop variants of biodegradable cross-linked composite hydrogel based on natural polypeptides (gelatin), polysaccharides components (chondroitin-4-sulphate and hyaluronic acid), in a ratio of 2:0.08:0.02 (w/w/w) and mixed with Si-hydroxyapatite (Si-Hap), in two ratios of 1:1 and 2:1 (w/w). Si-Hap was synthesized and characterized as a better alternative to conventional Hap. Subsequently, both composite hydrogel variants were cross-linked with (N, N-(3-dimethylaminopropyl)-N-ethyl carbodiimide (EDC) and enriched with a small bioactive molecule (icariin). The small molecule icariin (Ica) (C33H40O15) is the main active constituent (flavonoid) of Herba epimedium used in traditional Chinese medicine to cure bone- and cartilage-related disorders. Ica enhances osteogenic and chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), facilitates matrix calcification and increases the specific extracellular matrix (ECM) components synthesis by chondrocytes. Afterward, the composite hydrogels were characterized for their physicochemical properties in terms of the enzymatic biodegradation in the presence of type I collagenase and trypsin, the swelling capacity and the degree of crosslinking (TNBS assay). The cumulative release of Ica and real-time concentration were quantified at predetermined periods of time, according to the standard curve of standard Ica, after hydrogels incubation in saline buffer at physiological parameters. The obtained cross-linked composite hydrogels enriched with small-molecule Ica were also characterized for morphology by scanning electron microscopy (SEM). Their cytocompatibility was evaluated according to EN ISO 10993-5:2009 standard for medical device testing. Thus, analyses regarding cell viability (Live/Dead assay), cell proliferation (Neutral Red assay) and cell adhesion to composite hydrogels (SEM) were performed using NCTC clone L929 cell line. The final results showed that both cross-linked composite hydrogel variants enriched with Ica presented optimal physicochemical, structural and biological properties to be used as a natural scaffold able to repair osteochondral defects. The data did not reveal any toxicity of composite hydrogels in NCTC stabilized cell lines within the tested range of concentrations. Moreover, cells were capable of spreading and proliferating on both composite hydrogel surfaces. In conclusion, the designed biodegradable cross-linked composites enriched with Si and Ica are recommended for further testing as natural temporary scaffolds, which can allow cell migration and synthesis of new extracellular matrix within osteochondral defects.

Keywords: composites, gelatin, osteochondral defect, small molecule

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37 Multiple Plant-Based Cell Suspension as a Bio-Ink for 3D Bioprinting Applications in Food Technology

Authors: Yusuf Hesham Mohamed

Abstract:

Introduction: Three-dimensional printing technology includes multiple procedures that fabricate three-dimensional objects through consecutively layering two-dimensional cross-sections on top of each other. 3D bioprinting is a promising field of 3D printing, which fabricates tissues and organs by accurately controlling the proper arrangement of diverse biological components. 3D bioprinting uses software and prints biological materials and their supporting components layer-by-layer on a substrate or in a tissue culture plate to produce complex live tissues and organs. 3D food printing is an emerging field of 3D bioprinting in which the 3D printed products are food products that are cheap, require less effort to produce, and have more desirable traits. The Aim of the Study is the development of an affordable 3D bioprinter by altering a locally made CNC instrument with an open-source platform to suit the 3D bio-printer purposes. Later, we went through applying the prototype in several applications regarding food technology and drug testing, including the organ-On-Chip. Materials and Methods: An off-the-shelf 3D printer was modified by designing and fabricating the syringe unit, which was designed on the basis of the Milli-fluidics system. Sodium alginate and gelatin hydrogels were prepared, followed by leaf cell suspension preparation from narrow sections of Fragaria’s viable leaves. The desired 3D structure was modeled, and 3D printing preparations took place. Cell-free and cell-laden hydrogels were printed at room temperature under sterile conditions. Post printing curing process was performed. The printed structure was further studied. Results: Positive results have been achieved using the altered 3D bioprinter where a 3D hydrogel construct of two layers made of the combination of sodium alginate to gelatin (15%: 0.5%) has been printed. DLP 3D printer was used to design the syringe component with a transparent PLA-Pro resin for the creation of a microfluidics system having two channels altered to the double extruder. The hydrogel extruder’s design was based on peristaltic pumps, which utilized a stepper motor. The design and fabrication were made using DIY-3D printed parts. Hard plastic PLA was the material utilized for printing. SEM was used to carry out the porous 3D construct imaging. Multiple physical and chemical tests were performed in order to ensure that the cell line was suitable for hosting. Fragaria plant was developed by suspending Fragaria’s cells from its leaves using the 3D bioprinter. Conclusion: 3D bioprinting is considered to be an emerging scientific field that can facilitate and improve many scientific tests and studies. Thus, having a 3D bioprinter in labs is considered to be an essential requirement. 3D bioprinters are very expensive; however, the fabrication of a 3D printer into a 3D bioprinter can lower the cost of the bioprinter. The 3D bioprinter implemented made use of peristaltic pumps instead of syringe-based pumps in order to extend the ability to print multiple types of materials and cells.

Keywords: scaffold, eco on chip, 3D bioprinter, DLP printer

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36 Investigation the Effect of Velocity Inlet and Carrying Fluid on the Flow inside Coronary Artery

Authors: Mohammadreza Nezamirad, Nasim Sabetpour, Azadeh Yazdi, Amirmasoud Hamedi

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In this study OpenFOAM 4.4.2 was used to investigate flow inside the coronary artery of the heart. This step is the first step of our future project, which is to include conjugate heat transfer of the heart with three main coronary arteries. Three different velocities were used as inlet boundary conditions to see the effect of velocity increase on velocity, pressure, and wall shear of the coronary artery. Also, three different fluids, namely the University of Wisconsin solution, gelatin, and blood was used to investigate the effect of different fluids on flow inside the coronary artery. A code based on Reynolds Stress Navier Stokes (RANS) equations was written and implemented with the real boundary condition that was calculated based on MRI images. In order to improve the accuracy of the current numerical scheme, hex dominant mesh is utilized. When the inlet velocity increases to 0.5 m/s, velocity, wall shear stress, and pressure increase at the narrower parts.

Keywords: CFD, simulation, OpenFOAM, heart

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35 Ectopic Osteoinduction of Porous Composite Scaffolds Reinforced with Graphene Oxide and Hydroxyapatite Gradient Density

Authors: G. M. Vlasceanu, H. Iovu, E. Vasile, M. Ionita

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Herein, the synthesis and characterization of chitosan-gelatin highly porous scaffold reinforced with graphene oxide, and hydroxyapatite (HAp), crosslinked with genipin was targeted. In tissue engineering, chitosan and gelatin are two of the most robust biopolymers with wide applicability due to intrinsic biocompatibility, biodegradability, low antigenicity properties, affordability, and ease of processing. HAp, per its exceptional activity in tuning cell-matrix interactions, is acknowledged for its capability of sustaining cellular proliferation by promoting bone-like native micro-media for cell adjustment. Genipin is regarded as a top class cross-linker, while graphene oxide (GO) is viewed as one of the most performant and versatile fillers. The composites with natural bone HAp/biopolymer ratio were obtained by cascading sonochemical treatments, followed by uncomplicated casting methods and by freeze-drying. Their structure was characterized by Fourier Transform Infrared Spectroscopy and X-ray Diffraction, while overall morphology was investigated by Scanning Electron Microscopy (SEM) and micro-Computer Tomography (µ-CT). Ensuing that, in vitro enzyme degradation was performed to detect the most promising compositions for the development of in vivo assays. Suitable GO dispersion was ascertained within the biopolymer mix as nanolayers specific signals lack in both FTIR and XRD spectra, and the specific spectral features of the polymers persisted with GO load enhancement. Overall, correlations between the GO induced material structuration, crystallinity variations, and chemical interaction of the compounds can be correlated with the physical features and bioactivity of each composite formulation. Moreover, the HAp distribution within follows an auspicious density gradient tuned for hybrid osseous/cartilage matter architectures, which were mirrored in the mice model tests. Hence, the synthesis route of a natural polymer blend/hydroxyapatite-graphene oxide composite material is anticipated to emerge as influential formulation in bone tissue engineering. Acknowledgement: This work was supported by the project 'Work-based learning systems using entrepreneurship grants for doctoral and post-doctoral students' (Sisteme de invatare bazate pe munca prin burse antreprenor pentru doctoranzi si postdoctoranzi) - SIMBA, SMIS code 124705 and by a grant of the National Authority for Scientific Research and Innovation, Operational Program Competitiveness Axis 1 - Section E, Program co-financed from European Regional Development Fund 'Investments for your future' under the project number 154/25.11.2016, P_37_221/2015. The nano-CT experiments were possible due to European Regional Development Fund through Competitiveness Operational Program 2014-2020, Priority axis 1, ID P_36_611, MySMIS code 107066, INOVABIOMED.

Keywords: biopolymer blend, ectopic osteoinduction, graphene oxide composite, hydroxyapatite

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34 Fabrication of Antimicrobial Dental Model Using Digital Light Processing (DLP) Integrated with 3D-Bioprinting Technology

Authors: Rana Mohamed, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Bio-fabrication is a multidisciplinary research field that combines several principles, fabrication techniques, and protocols from different fields. The open-source-software movement is a movement that supports the use of open-source licenses for some or all software as part of the broader notion of open collaboration. Additive manufacturing is the concept of 3D printing, where it is a manufacturing method through adding layer-by-layer using computer-aided designs (CAD). There are several types of AM system used, and they can be categorized by the type of process used. One of these AM technologies is Digital light processing (DLP) which is a 3D printing technology used to rapidly cure a photopolymer resin to create hard scaffolds. DLP uses a projected light source to cure (Harden or crosslinking) the entire layer at once. Current applications of DLP are focused on dental and medical applications. Other developments have been made in this field, leading to the revolutionary field 3D bioprinting. The open-source movement was started to spread the concept of open-source software to provide software or hardware that is cheaper, reliable, and has better quality. Objective: Modification of desktop 3D printer into 3D bio-printer and the integration of DLP technology and bio-fabrication to produce an antibacterial dental model. Method: Modification of a desktop 3D printer into a 3D bioprinter. Gelatin hydrogel and sodium alginate hydrogel were prepared with different concentrations. Rhizome of Zingiber officinale, Flower buds of Syzygium aromaticum, and Bulbs of Allium sativum were extracted, and extractions were selected on different levels (Powder, aqueous extracts, total oils, and Essential oils) prepared for antibacterial bioactivity. Agar well diffusion method along with the E. coli have been used to perform the sensitivity test for the antibacterial activity of the extracts acquired by Zingiber officinale, Syzygium aromaticum, and Allium sativum. Lastly, DLP printing was performed to produce several dental models with the natural extracted combined with hydrogel to represent and simulate the Hard and Soft tissues. Result: The desktop 3D printer was modified into 3D bioprinter using open-source software Marline and modified custom-made 3D printed parts. Sodium alginate hydrogel and gelatin hydrogel were prepared at 5% (w/v), 10% (w/v), and 15%(w/v). Resin integration with the natural extracts of Rhizome of Zingiber officinale, Flower buds of Syzygium aromaticum, and Bulbs of Allium sativum was done following the percentage 1- 3% for each extract. Finally, the Antimicrobial dental model was printed; exhibits the antimicrobial activity, followed by merging with sodium alginate hydrogel. Conclusion: The open-source movement was successful in modifying and producing a low-cost Desktop 3D Bioprinter showing the potential of further enhancement in such scope. Additionally, the potential of integrating the DLP technology with bioprinting is a promising step toward the usage of the antimicrobial activity using natural products.

Keywords: 3D printing, 3D bio-printing, DLP, hydrogel, antibacterial activity, zingiber officinale, syzygium aromaticum, allium sativum, panax ginseng, dental applications

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33 Treatment of Neuronal Defects by Bone Marrow Stem Cells Differentiation to Neuronal Cells Cultured on Gelatin-PLGA Scaffolds Coated with Nano-Particles

Authors: Alireza Shams, Ali Zamanian, Atefehe Shamosi, Farnaz Ghorbani

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Introduction: Although the application of a new strategy remains a remarkable challenge for treatment of disabilities due to neuronal defects, progress in Nanomedicine and tissue engineering, suggesting the new medical methods. One of the promising strategies for reconstruction and regeneration of nervous tissue is replacing of lost or damaged cells by specific scaffolds after Compressive, ischemic and traumatic injuries of central nervous system. Furthermore, ultrastructure, composition, and arrangement of tissue scaffolds are effective on cell grafts. We followed implantation and differentiation of mesenchyme stem cells to neural cells on Gelatin Polylactic-co-glycolic acid (PLGA) scaffolds coated with iron nanoparticles. The aim of this study was to evaluate the capability of stem cells to differentiate into motor neuron-like cells under topographical cues and morphogenic factors. Methods and Materials: Bone marrow mesenchymal stem cells (BMMSCs) was obtained by primary cell culturing of adult rat bone marrow got from femur bone by flushing method. BMMSCs were incubated with DMEM/F12 (Gibco), 15% FBS and 100 U/ml pen/strep as media. Then, BMMSCs seeded on Gel/PLGA scaffolds and tissue culture (TCP) polystyrene embedded and incorporated by Fe Nano particles (FeNPs) (Fe3o4 oxide (M w= 270.30 gr/mol.). For neuronal differentiation, 2×10 5 BMMSCs were seeded on Gel/PLGA/FeNPs scaffolds was cultured for 7 days and 0.5 µ mol. Retinoic acid, 100 µ mol. Ascorbic acid,10 ng/ml. Basic fibroblast growth factor (Sigma, USA), 250 μM Iso butyl methyl xanthine, 100 μM 2-mercaptoethanol, and 0.2 % B27 (Invitrogen, USA) added to media. Proliferation of BMMSCs was assessed by using MTT assay for cell survival. The morphology of BMMSCs and scaffolds was investigated by scanning electron microscopy analysis. Expression of neuron-specific markers was studied by immunohistochemistry method. Data were analyzed by analysis of variance, and statistical significance was determined by Turkey’s test. Results: Our results revealed that differentiation and survival of BMMSCs into motor neuron-like cells on Gel/PLGA/FeNPs as a biocompatible and biodegradable scaffolds were better than those cultured in Gel/PLGA in absence of FeNPs and TCP scaffolds. FeNPs had raised physical power but decreased capacity absorption of scaffolds. Well defined oriented pores in scaffolds due to FeNPs may activate differentiation and synchronized cells as a mechanoreceptor. Induction effects of magnetic FeNPs by One way flow of channels in scaffolds help to lead the cells and can facilitate direction of their growth processes. Discussion: Progression of biological properties of BMMSCs and the effects of FeNPs spreading under magnetic field was evaluated in this investigation. In vitro study showed that the Gel/PLGA/FeNPs scaffold provided a suitable structure for motor neuron-like cells differentiation. This could be a promising candidate for enhancing repair and regeneration in neural defects. Dynamic and static magnetic field for inducing and construction of cells can provide better results for further experimental studies.

Keywords: differentiation, mesenchymal stem cells, nano particles, neuronal defects, Scaffolds

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32 Modeling of a Pendulum Test Including Skin and Muscles under Compression

Authors: M. J. Kang, Y. N. Jo, H. H. Yoo

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Pendulum tests were used to identify a stretch reflex and diagnose spasticity. Some researches tried to make a mathematical model to simulate the motions. Thighs are subject to compressive forces due to gravity during a pendulum test. Therefore, it affects knee trajectories. However, the most studies on the pendulum tests did not consider that conditions. We used Kelvin-Voight model as compression model of skin and muscles. In this study, we investigated viscoelastic behaviors of skin and muscles using gelatin blocks from experiments of the vibration of the compliantly supported beam. Then we calculated a dynamic stiffness and loss factors from the experiment and estimated a damping coefficient of the model. We also did pendulum tests of human lower limbs to validate the stiffness and damping coefficient of a skin model. To simulate the pendulum motion, we derive equations of motion. We used stretch reflex activation model to estimate muscle forces induced by the stretch reflex. To validate the results, we compared the activation with electromyography signals during experiments. The compression behavior of skin and muscles in this study can be applied to analyze sitting posture as wee as developing surgical techniques.

Keywords: Kelvin-Voight model, pendulum test, skin and muscles under compression, stretch reflex

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31 The Study on Blast Effect of Polymer Gel by Trazul Lead Block Test and Concrete Block Test

Authors: Young-Hun Ko, Seung-Jun Kim, Khaqan Baluch, Hyung- Sik Yang

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In this study, the polymer gel was used as coupling material in a blasting hole and its comparison was made with other coupling materials like sand, water, and air. Trazul lead block test and AUTODYN numerical analysis were conducted to analyze the effects of the coupling materials on the intensity of the explosion, as well as the verification tests were conducted by using concrete block test. The emulsion explosives were used in decoupling conditions, sand, water, and polymer gel were used as the coupling materials. The lead block test and the numerical analysis showed that the expansion of the blast hole in the lead block was similar to that of the water and gelatin and followed by sand and air conditions. The validation of concrete block test result showed the similar result as Trazul lead block test and the explosion strength was measured at 0.8 for polymer gel, 0.7 for sand, and 0.6 for no coupling material, in comparison to the full charge (1.0) case.

Keywords: Trazul lead block test, AUTODYN numerical analysis, coupling material, polymer gel, soil covering concrete block explosion test

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30 Biochemical and Molecular Analysis of Staphylococcus aureus Various Isolates from Different Places

Authors: Kiran Fatima, Kashif Ali

Abstract:

Staphylococcus aureus is an opportunistic human as well as animal pathogen that causes a variety of diseases. A total of 70 staphylococci isolates were obtained from soil, water, yogurt, and clinical samples. The likely staphylococci clinical isolates were identified phenotypically by different biochemical tests. Molecular identification was done by PCR using species-specific 16S rRNA primer pairs, and finally, 50 isolates were found to be positive as Staphylococcus aureus, sciuri, xylous and cohnii. Screened isolates were further analyzed by several microbiological diagnostics tests, including gram staining, coagulase, capsule, hemolysis, fermentation of glucose, lactose, maltose, and sucrose tests enzymatic reactions. It was found that 78%, 81%, and 51% of isolates were positive for gelatin hydrolysis, protease, and lipase activities, respectively. Antibiogram analysis of isolated Staphylococcus aureus strains with respect to different antimicrobial agents revealed resistance patterns ranging from 57 to 96%. Our study also shows 70% of strains to be MRSA, 54.3% as VRSA, and 54.3% as both MRSA and VRSA. All the identified isolates were subjected to detection of mecA, nuc, and hlb genes, and 70%, 84%, and 40% were found to harbour mecA, nuc, and hlb genes, respectively. The current investigation is highly important and informative for the high-level multidrug-resistant Staphylococcus aureus infections inclusive also of methicillin and vancomycin.

Keywords: MRSA, VRSA, mecA, MSSA

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29 Microencapsulation of Probiotic and Evaluation for Viability, Antimicrobial Property and Cytotoxic Activities of its Postbiotic Metabolites on MCF-7 Breast Cancer Cell Line

Authors: Nkechi V. Enwuru, Bullum Nkeki, Elizabeth A. Adekoya, Olumide A. Adebesin, Rebecca F. Peters, Victoria A. Aikhomu, Mendie E. U.

Abstract:

Background: Probiotics are live microbial feed supplement beneficial for host. Probiotics and their postbiotic products have been used to prevent or treat various health conditions. However, the products cell viability is often low due to harsh conditions subjected during processing, handling, storage, and gastrointestinal transit. These strongly influence probiotics’ benefits; thus, viability is essential for probiotics to produce health benefits for the host. Microencapsulation is a promising technique with considerable effects on probiotic survival. The study is aimed to formulate a microencapsulated probiotic and evaluate its viability, antimicrobial efficacy, and cytotoxic activity of its postbiotic on the MCF-7 breast cancer cell line. Method: Human and animal raw milk were sampled for lactic acid bacteria. The isolated bacteria were identified using conventional and VITEK 2 systems. The identified lactic acid bacterium was encapsulated using spray-dried and extrusion methods. The free, encapsulated, and chitosan-coated encapsulated probiotics were tested for viability in simulated-gastric intestinal (SGI) fluid and different storage conditions at refrigerated (4oC) and room (25oC) temperatures. The disintegration time and weight uniformity of the spray-dried hard gelatin capsules were tested. The antimicrobial property of free and encapsulated probiotics was tested against enteric pathogenic isolates from antiretroviral therapy (ART) treated HIV-positive patients. The postbiotic of the free cells was extracted, and its cytotoxic effect on the MCF-7 breast cancer cell line was tested through an MTT assay. Result: The Lactobacillus plantarum was isolated from animal raw milk. Zero-size hard gelatin L. plantarum capsules with granules within a size range of 0.71–1.00 mm diameter was formulated. The disintegration time ranges from 2.14±0.045 to 2.91±0.293 minutes, while the average weight is 502.1mg. Simulated gastric solution significantly affected viability of both free and microcapsules. However, the encapsulated cells were more protected and viable due to impermeability in the microcapsules. Furthermore, the viability of free cells stored at 4oC and 25oC were less than 4 log CFU/g and 6 log CFU/g respectively after 12 weeks. However, the microcapsules stored at 4oC achieved the highest viability among the free and microcapsules stored at 25oC and the free cells stored at 4oC. Encapsulated cells were released in the simulated gastric fluid, viable and effective against the enteric pathogens tested. However, chitosan-coated calcium alginate encapsulated probiotics significantly inhibited Shigella flexneri, Candida albicans, and Escherichia coli. The Postbiotic Metabolites (PM) of L. plantarum produced a cytotoxic effect on the MCF-7 breast cancer cell line. The postbiotic showed significant cytotoxic activity similar to 5FU, a standard antineoplastic agent. The inhibition concentration of 50% growth (IC50) of postbiotic metabolite K3 is low and consistent with the IC50 of the positive control (Cisplatin). Conclusions: Lactobacillus plantarum postbiotic exhibited a cytotoxic effect on the MCF-7 breast cancer cell line and could be used as combined adjuvant therapy in breast cancer management. The microencapsulation technique protects the probiotics, improving their viability and delivery to the gastrointestinal tract. Chitosan enhances antibacterial efficacy; thus, chitosan-coated microencapsulated L. plantarum probiotics could be more effective and used as a combined therapy in HIV management of opportunistic enteric infection.

Keywords: probiotics, encapsulation, gastrointestinal conditions, antimicrobial effect, postbiotic, cytotoxicity effect

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28 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

Abstract:

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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27 Tofu Flour as a Protein Sources

Authors: Dicky Eka Putra, S. P. Nadia Chairunissa, Lidia Paramita, Roza Hartati, Ice Yolanda Puri

Abstract:

Background: Soy bean and the products such as tofu, tempeh and soy milk are famous in the community. Moreover, another product is tofu flour which is not familiar in Indonesia yet and it is well known as Okara. There are massive differences of energy, protein and carbohydrate between them which is know as good for protein sources as well. Unfortunately, it is seldom used as food variety. Basically, it can be benefit in order to create many products for example cakes, snacks and some desserts. Aim: the study was in order to promote the benefit of tofu flour as school feeding of elementary school and baby porridge and also to compare the nutrient. Method: Soy pulp was filtered and steamed approximately 30 minutes. Then, it was put at a plate under sunrise or barked on the oven for 10 hours at 800C. When it have dried and milling and tofu flour is ready to be used. Result: Tofu flour could be used as substitute of flour and rice flour when people want to cook some foods. In addition, some references said that soy bean is good for a specific remedy for the proper functioning of the heart, liver, kidneys, stomach, and bowels, constipation, as a stimulant for the lungs, for eradication of poison from the system, improving the complexion by cleaning the skin of impurities, and stimulating the growth and appearance of the hair. Discussion: Comparing between soy bean, tofu and tofu flour which has difference amount of nutrients. For example energy 382 kcal, 79 kcal and 393 kcal respectively and also protein 30.2 kcal, 7.8 kcal, and 17.4 kcal. In addition, carbohydrate of soy pulp was high than soy bean and tofu (30.1 kcal). Finally, local should replace flour, rice and gelatin rice flour with tofu flour.

Keywords: tofu flour, protein, soy bean, school feeding

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26 A 3D Bioprinting System for Engineering Cell-Embedded Hydrogels by Digital Light Processing

Authors: Jimmy Jiun-Ming Su, Yuan-Min Lin

Abstract:

Bioprinting has been applied to produce 3D cellular constructs for tissue engineering. Microextrusion printing is the most common used method. However, printing low viscosity bioink is a challenge for this method. Herein, we developed a new 3D printing system to fabricate cell-laden hydrogels via a DLP-based projector. The bioprinter is assembled from affordable equipment including a stepper motor, screw, LED-based DLP projector, open source computer hardware and software. The system can use low viscosity and photo-polymerized bioink to fabricate 3D tissue mimics in a layer-by-layer manner. In this study, we used gelatin methylacrylate (GelMA) as bioink for stem cell encapsulation. In order to reinforce the printed construct, surface modified hydroxyapatite has been added in the bioink. We demonstrated the silanization of hydroxyapatite could improve the crosslinking between the interface of hydroxyapatite and GelMA. The results showed that the incorporation of silanized hydroxyapatite into the bioink had an enhancing effect on the mechanical properties of printed hydrogel, in addition, the hydrogel had low cytotoxicity and promoted the differentiation of embedded human bone marrow stem cells (hBMSCs) and retinal pigment epithelium (RPE) cells. Moreover, this bioprinting system has the ability to generate microchannels inside the engineered tissues to facilitate diffusion of nutrients. We believe this 3D bioprinting system has potential to fabricate various tissues for clinical applications and regenerative medicine in the future.

Keywords: bioprinting, cell encapsulation, digital light processing, GelMA hydrogel

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25 Preparation and In vitro Characterization of Nanoparticle Hydrogel for Wound Healing

Authors: Rajni Kant Panik

Abstract:

The aim of the present study was to develop and evaluate mupirocin loaded nanoparticle incorporated into hydrogel as an infected wound healer. Incorporated Nanoparticle in hydrogel provides a barrier that effectively prevents the contamination of the wound and further progression of infection to deeper tissues. Hydrogel creates moist healing environment on wound space with good fluid absorbance. Nanoparticles were prepared by double emulsion solvent evaporation method using different ratios of PLGA polymer and the hydrogels was developed using sodium alginate and gelatin. Further prepared nanoparticles were then incorporated into the hydrogels. The formulations were characterized by FT-IR and DSC for drug and polymer compatibility and surface morphology was studied by TEM. Nanoparticle hydrogel were evaluated for their size, shape, encapsulation efficiency and for in vitro studies. The FT-IR and DSC confirmed the absence of any drug polymer interaction. The average size of Nanoparticle was found to be in range of 208.21-412.33 nm and shape was found to be spherical. The maximum encapsulation efficiency was found to be 69.03%. The in vitro release profile of Nanoparticle incorporated hydrogel formulation was found to give sustained release of drug. Antimicrobial activity testing confirmed that encapsulated drug preserve its effectiveness. The stability study confirmed that the formulation prepared were stable. Present study complements our finding that mupirocin loaded Nanoparticle incorporated into hydrogel has the potential to be an effective and safe novel addition for the release of mupirocin in sustained manner, which may be a better option for the management of wound. These finding also supports the progression of antibiotic via hydrogel delivery system is a novel topical dosage form for the management of wound.

Keywords: hydrogel, nanoparticle, PLGA, wound healing

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24 Development and Characterization of Controlled Release Photo Cross-Linked Implants for Ocular Delivery of Triamcinolone Acetonide

Authors: Ravi Sheshala, Annie Lee, Ai Lin Ong, Ling Ling Cheu, Thiagarajan Madheswaran, Thankur R. R. Singh

Abstract:

The objectives of the present research work were to develop and characterize biodegradable controlled release photo cross-linked implants of Triamcinolone Acetonide (TA) for the treatment of chronic ocular diseases. The photo cross-linked implants were prepared using film casting technique by mixing TA (2.5%) polyethylene glycol diacrylate (PEGDA 700), pore formers (mannitol, maltose, and gelatin) and the photoinitiator (Irgacure 2959). The resulting mixture was injected into moulds using 21 G and subjected to photocrosslinking at 365 nm. Scanning electron microscopy results demonstrated that more pores were formed in the films with the increase in the concentration of pore formers from 2%-10%. The maximum force required to break the films containing 2-10% of pore formers were determined in both dry and wet conditions using texture analyzer and found that films in a dry condition required a higher force to break compared to wet condition and blank films. In vitro drug release from photo cross-linked films were determined by incubating samples in 50 ml PBS pH 7.4 at 37 C and the samples were analyzed for drug release by HPLC. The films demonstrated a biphasic release profile i.e. an initial burst release (<20%) on the first day followed by a constant and continuous drug release in a controlled manner for 42 days. The drug release from all formulations followed the first-order release pattern and the combination of diffusion and erosion release mechanism. In conclusion, the developed formulations were able to provide controlled drug delivery to treat the chronic ocular diseases.

Keywords: controlled release, ophthalmic, PEGDA, photocrosslinking, pore formers

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23 Partial Purification and Characterization of a Low Molecular Weight and Industrially Important Chitinase and a Chitin Deacetylase Enzyme from Streptomyces Chilikensis RC1830, a Novel Strain Isolated from Chilika Lake, India

Authors: Lopamudra Ray, Malla Padma, Dibya Bhol, Samir Ranjan Mishra, A. N. Panda, Gurdeep Rastogi, T. K. Adhya, Ajit Kumar Pattnaik, Mrutyunjay Suar, Vishakha Raina

Abstract:

Chilika Lake is the largest coastal estuarine brackish water lagoon in Asia situated on the east coast of India and is a designated Ramsar site. In the current study, several chitinolytic microorganisms were isolated and screened by appearance of clearance zone on 0.5% colloidal chitin agar plate. A strain designated as RC 1830 displayed maximum colloidal chitin degradation by release of 112 μmol/ml/min of N-acetyl D-glucosamine (GlcNAc) in 48h. The strain was taxonomically identified by polyphasic approach based on a range of phenotypic and genotypic properties and was found to be a novel species named Streptomyces chilikensis RC1830. The organism was halophilic (12% NaCl w/v), alkalophilic (pH10) and was capable of hydrolyzing chitin, starch, cellulose, gelatin, casein, tributyrin and tween 80. The partial purification of chitinase enzymes from RC1830 was performed by DEAE Sephacel anion exchange chromatography which revealed the presence of a very low molecular weight chitinase(10.5kD) which may be a probable chitobiosidase enzyme. The study reports the presence of a low MW chitinase (10.5kD) and a chitin decaetylase from a novel Streptomyces strain RC1830 isolated from Chilika Lake. Previously chitinases less than 20.5kD have not been reported from any other Streptomyces species. The enzymes was characterized with respect to optimum pH, temperature, and substrate specificity and temperature stability.

Keywords: chitinases, chitobiosidase, Chilika Lake, India

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22 The Influence of Amygdalin on Glioblastoma Multiforme Cell Lines

Authors: Sylwia K. Naliwajko, Justyna Moskwa, Patryk Nowakowski, Renata Markiewicz-Zukowska, Krystyna Gromkowska-Kepka, Anna Puscion-Jakubik, Maria H. Borawska

Abstract:

Amygdalin is found in many fruit seeds, including apricot, peach, quince, apples, and almonds. Amygdalin (also named vitamin B17), as well as its sources, are commonly used as an alternative therapy or prevention of cancer. The potential activity of amygdalin is related to its enzymatic degradation to the hydrogen cyanide. Hydrogen cyanide is a toxic substance that causes liver and nerves damage, fever, coma or even death. Amygdalin is much better tolerated after intravenous than oral administration. The aim of this study was to examine the influence of amygdalin on glioblastoma multiforme cell lines. Three glioblastoma multiforme cell lines – U87MG, T98, LN18 were incubated (48 h) with amygdalin in concentrations 100, 250, 500, 1000 and 2000 µg/mL. The MTT (Thiazolyl Blue Tetrazolium Bromide) test and DNA binding test by [3H]-thymidine incorporation were used to determine the anti-proliferative activity of amygdalin. The secretion of metalloproteinases (MMP2 and MMP-9) from U87MG cells was estimated by gelatin zymography. The statistical analysis was performed using Statistica v. 13.0 software. The data was presented as a % of control. Amygdalin did not show significant inhibition of viability of all the glioblastoma cells in concentrations 100, 250, 500, 1000 µg/mL. In 2000 µg/mL there were significant differences compared to the control, but inhibition of viability was less than 20% (more than 80% of control). The average viability of U87MG cells was 92,0±4%, T98G: 85,8±3% and LN18: 94,7±2% of the control. There was no dose-response viability, and IC50 value was not recognized. DNA binding in U87MG cells was not inhibited (109,0±3 % of control). After treatment with amygdalin, we observed significantly increased secretion of MMP2 and MMP9 in U87MG cells (130,3±14% and 112,0±5% of control, respectively). Our results suggest that amygdalin has no anticancer activity in glioblastoma cell lines.

Keywords: amygdalin, anticancer, cell line, glioblastoma

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21 Incorporation of Growth Factors onto Hydrogels via Peptide Mediated Binding for Development of Vascular Networks

Authors: Katie Kilgour, Brendan Turner, Carly Catella, Michael Daniele, Stefano Menegatti

Abstract:

In vivo, the extracellular matrix (ECM) provides biochemical and mechanical properties that are instructional to resident cells to form complex tissues with characteristics to develop and support vascular networks. In vitro, the development of vascular networks can be guided by biochemical patterning of substrates via spatial distribution and display of peptides and growth factors to prompt cell adhesion, differentiation, and proliferation. We have developed a technique utilizing peptide ligands that specifically bind vascular endothelial growth factor (VEGF), erythropoietin (EPO), or angiopoietin-1 (ANG1) to spatiotemporally distribute growth factors to cells. This allows for the controlled release of each growth factor, ultimately enhancing the formation of a vascular network. Our engineered tissue constructs (ETCs) are fabricated out of gelatin methacryloyl (GelMA), which is an ideal substrate for tailored stiffness and bio-functionality, and covalently patterned with growth factor specific peptides. These peptides mimic growth factor receptors, facilitating the non-covalent binding of the growth factors to the ETC, allowing for facile uptake by the cells. We have demonstrated in the absence of cells the binding affinity of VEGF, EPO, and ANG1 to their respective peptides and the ability for each to be patterned onto a GelMA substrate. The ability to organize growth factors on an ETC provides different functionality to develop organized vascular networks. Our results demonstrated a method to incorporate biochemical cues into ETCs that enable spatial and temporal control of growth factors. Future efforts will investigate the cellular response by evaluating gene expression, quantifying angiogenic activity, and measuring the speed of growth factor consumption.

Keywords: growth factor, hydrogel, peptide, angiogenesis, vascular, patterning

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20 Myomectomy and Blood Loss: A Quality Improvement Project

Authors: Ena Arora, Rong Fan, Aleksandr Fuks, Kolawole Felix Akinnawonu

Abstract:

Introduction: Leiomyomas are benign tumors that are derived from the overgrowth of uterine smooth muscle cells. Women with symptomatic leiomyomas who desire future fertility, myomectomy should be the standard surgical treatment. Perioperative hemorrhage is a common complication in myomectomy. We performed the study to investigate blood transfusion rate in abdominal myomectomies, risk factors influencing blood loss and modalities to improve perioperative blood loss. Methods: Retrospective chart review was done for patients who underwent myomectomy from 2016 to 2022 at Queens hospital center, New York. We looked at preoperative patient demographics, clinical characteristics, intraoperative variables, and postoperative outcomes. Mann-Whitney U test were used for parametric and non-parametric continuous variable comparisons, respectively. Results: A total of 159 myomectomies were performed between 2016 and 2022, including 1 laparoscopic, 65 vaginal and 93 abdominal. 44 patients received blood transfusion during or within 72 hours of abdominal myomectomy. The blood transfusion rate was 47.3%. Blood transfusion rate was found to be twice higher than the average documented rate in literature which is 20%. Risk factors identified were black race, preoperative hematocrit<30%, preoperative blood transfusion within 72 hours, large fibroid burden, prolonged surgical time, and abdominal approach. Conclusion: Preoperative optimization with iron supplements or GnRH agonists is important for patients undergoing myomectomy. Interventions to decrease intra operative blood loss should include cell saver, tourniquet, vasopressin, misoprostol, tranexamic acid and gelatin-thrombin matrix hemostatic sealant.

Keywords: myomectomy, perioperative blood loss, cell saver, tranexamic acid

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19 Encapsulated Western Red Cedar (Thuja Plicata) Essential Oil as a Prospective Biopesticide against Phytophthora Pathogens

Authors: Aleksandar M. Radojković, Jovana M. Ćirković, Sanja Z. Perać, Jelena N. Jovanović, Zorica M. Branković, Slobodan D. Milanović, Ivan Lj. Milenković, Jovan N. Dobrosavljević, Nemanja V. Simović, Vanja M. Tadić, Ana R. Žugić, Goran O. Branković

Abstract:

In many parts of the world, various Phytophthora species pose a serious threat to forests and crops. With the rapidly growing international trade in plants and the ongoing impacts of climate change, the harmful effects of plant pathogens of the genus Phytophthora are increasing, damaging the biodiversity and sustainability of forest ecosystems. This genus is one of the most destructive plant pathogens, causing the majority of fine root (66%) and collar rot diseases (90%) of woody plant species worldwide. Eco-friendly biopesticides, based on plant-derived products, such as essential oils (EOs), are one of the promising solutions to this problem. In this study, among three different EOs investigated (Chamaecyparis lawsoniana (A. Murr.) Parl., Thuja plicata Donn ex D.Don and Juniperus communis L.), western red cedar (Thuja plicata) essential oil almost completely inhibited the growth of three Phytophthora species (P. plurivora Jung and Burgess, P. quercina Jung, and P. ×cambivora (Petri) Buisman) during seven days of exposure for the EO concentrations of 0.1% and 0.5% (v/v). To prolong the inhibiting effect, Thuja plicata EO was encapsulated into a biopolymer matrix consisting of a chitosan-gelatin mixture to form a water-in-oil emulsion. This approach allowed the prolonged effect of the essential oil by its slow release from the biopolymer matrix and protection of the active components from atmospheric influences. Thus, it was demonstrated that encapsulated Thuja plicata EO consisting of sustainable bioproducts is efficient in controlling of Phytophthora species and can be considered a means of protection in natural and semi-natural ecosystems.

Keywords: emulsions, essential oils, phytophthora, thuja plicata

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18 First Report of Rahnella Victoriana Associated with Walnut Decline

Authors: Mohammadreza Hajialigol, Nargues Falahi Charkhabi, Fatemeh Shahryari, Saadat Sarikhani

Abstract:

BACKGROUND AND OBJECTIVES Iran is the third producer of Persian walnut worldwide. However, its walnut trees have been under threat from decline during last decade. Walnut canker caused by B. nigrifluens and B. rubrifaciens was recorded in multiple regions of Iran. Furthermore, Brenneria rosae subsp. rosae and Gibbsiella quercinecans were recently recognized as responsible for walnut decline in northwestern Iran. This study aimed to identify the causal agent of walnut decline in Kermanshah and Isfahan. MATERIAL AND METHODS Symptomatic samples were collected from affected walnut trees of Kermanshah and Isfahan provinces. The pathogenicity of strains was proved on immature walnut fruits cv. ‘Hartley’ and young green twigs of two-year-old walnut seedling cv. ‘Chandler’. Pathogenic strains were subjected to conventional phenotypic tests. 16S rRNA, gyrB, and infB genes were partially amplified and sequenced. RESULTS Irregular longitudinal cankers and dark lesions were observed in the outer and inner bark, respectively. Twenty-four strains were isolated on EMB-agar media. Fourteen strains were able to cause necrosis and a dark-colored region in the mesocarp and on young green twigs around the inoculation site 14 and 30 days post-inoculation, respectively. Strains were able to hydrolyze Tween 20, Tween 80, gelatin and esculin, however, did not produce indole or urease. Pairwise comparison, the 16S rRNA gene nucleotide sequences of strain I2 were 100% identical with those of Rahnella victoriana FRB 225T. Moreover, a phylogenetic tree reconstructed based on the concatenated sequences of two housekeeping gene fragments, gyrB (601 bp) and infB (615 bp), revealed that the strains I2, I5, and KE6 were clustered with R. victoriana FRB 225T. CONCLUSION To the best of our knowledge, this is the first report of R. victoriana in association with walnut decline. This result is necessary to find resistant genotypes.

Keywords: emerging pathogens, Iran, juglans regia, MLSA

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