Search results for: tissue invasion

Authors: Hanjun Yun, Gun Bae, Nabin Paul, Cheolhee Moon

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Flexible OLED is an important technology for the next generation display over various kinds of applications. However, the organic materials of OLEDs degrade rapidly under the invasion of oxygen and water moisture. The degradation causes the formation of non-emitting areas which gradually suppress the device brightness, ultimately the lifetime of the device decreasing rapidly. Until now, the most suitable sealing process of the flexible OLED devices is a thin film encapsulation (TFE). However, TFE consists of a multilayer thin-film structure with organic-inorganic materials, so the cost is expensive and the process time is long. Another problem is that the blocking characteristics from the moisture and oxygen are not perfect. Therefore, the encapsulation of the flexible OLED device is a still key technical issue for the successful market entry. In this study, we are to introduce an auxiliary sealing line between the two flexible substrates. The electrode lines were formed on the substrates which have a SiNx barrier coating layer. To induce the solid phase diffusion process between the SiNx layer and the electrode lines, the electrode materials were determined as Al-Si composition. Thermal energy was supplied for both the SiNx layer and Al-Si electrode lines within the furnace to induce the interfacial bonding through the solid phase diffusion of Si. We printed a test pattern for the edge of the flexible PET substrate of 3cm*3cm size. Experimental conditions such as heating temperature, heating time were optimized to get enough adhesion strength which was estimated through the competitive bending test. Finally, OLED devices with flexible PET substrate of 3cm*3cm size were manufactured to investigate the blocking characteristics as an encapsulation layer.

Keywords: barrier, encapsulation, OLED, solid phase diffusion

Procedia PDF Downloads 237

Authors: Brian Chi Yan Cheng, Hui Guo, Tao Su, Xiu‐qiong Fu, Ting Li, Zhi‐ling Yu

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Rheumatoid arthritis (RA) affects around 1% of the globe population. Yet, there is still no cure for RA. Toll-like receptor 4 (TLR4) signalling has been found to be involved in the pathogenesis of RA, making it a potential therapeutic target for RA treatment. A herbal formula (RL) consisting of fruits of Rosa Multiflora (Eijitsu rose) and flowers of Lonicera Japonica (Japanese honeysuckle) has been used in treating various inflammatory disorders for more than a thousand year. Both of them are rich sources of nutrients and bioactive phytochemicals, which can be used in producing different food products and supplements. In this study, we would evaluate the anti-arthritic effect of RL on collagen-induced arthritis (CIA) in rats and investigate the involvement of TLR4 signaling in the mode of action of RL. Anti-arthritic efficacy was evaluated using CIA rats induced by bovine type II collagen. The treatment groups were treated with RL (82.5, 165, and 330 mg/kg bw per day, p.o.) or positive control indomethacin (0.25 mg/kg bw per day, p.o.) for 35 days. Clinical signs (hind paw volume and arthritis severity scores), changes in serum inflammatory mediators, pro-/antioxidant status, histological and radiographic changes of joints were investigated. Spleens and peritoneal macrophages were used to determine the effects of RL on innate and adaptive immune responses in CIA rats. The involvement of TLR4 signalling pathways in the anti-arthritic effect of RL was examined in cartilage tissue of CIA rats, murine RAW264.7 macrophages and human THP-1 monocytic cells. The severity of arthritis in the CIA rats was significantly attenuated by RL. Antioxidant status, histological score and radiographic score were efficiently improved by RL. RL could also dose-dependently inhibit pro-inflammatory cytokines in serum of CIA rats. RL significantly inhibited the production of various pro-inflammatory mediators, the expression and/or activity of the components of TLR4 signalling pathways in animal tissue and cell lines. RL possesses anti-arthritic effect on collagen-induced arthritis in rats. The therapeutic effect of RL may be related to its inhibition on pro-inflammatory cytokines in serum. The inhibition of the TAK1/NF-κB and TAK1/MAPK pathways participate in the anti-arthritic effects of RL. This provides a pharmacological justification for the dietary use of RL in the control of various arthritic diseases. Further investigation should be done to develop RL into a anti-arthritic food products and/or supplements.

Keywords: japanese honeysuckle, rheumatoid arthritis, rosa multiflora, rosehip

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Authors: Tuji Mohammed Sani

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Back ground: An open tibial fracture is an injury where the fractured bone directly communicates with the outside environment. Due to the specific anatomical features of the tibia (limited soft tissue coverage), more than quarter of its fractures are classified as open, representing the most common open long-bone injuries. Open tibial fractures frequently cause significant bone comminution, periosteal stripping, soft tissue loss, contamination and are prone to bacterial entry with biofilm formation, which increases the risk of deep bone infection. Objective: The main objective of the study was to determine Prevalence of infection and its associated factors in surgically treated open tibial fracture in Addis Ababa Burn Emergency and Trauma (AaBET) center. Method: A facility based retrospective cross-sectional study was conducted among patient treated for open tibial fracture at AaBET center from September 2018 to September 2021. The data was collected from patient’s chart using structured data collection form, and Data was entered and analyzed using SPSS version 26. Bivariable and multiple binary logistic regression were fitted. Multicollinearity was checked among candidate variables using variance inflation factor and tolerance, which were less than 5 and greater than 0.2, respectively. Model adequacy were tested using Hosmer-Lemeshow goodness of fitness test (P=0.711). AOR at 95% CI was reported, and P-value < 0.05 was considered statistically significant. Result: This study found that 33.9% of the study participants had an infection. Initial IV antibiotic time (AOR=2.924, 95% CI:1.160- 7.370) and time of wound closure from injury (AOR=3.524, 95% CI: 1.798-6.908), injury to admission time (AOR=2.895, 95% CI: 1.402 – 5.977). and definitive fixation method (AOR=0.244, 95% CI: 0.113 – 0.4508) were the factors found to have a statistically significant association with the occurrence of infection. Conclusion: The rate of infection in open tibial fractures indicates that there is a need to improve the management of open tibial fracture treated at AaBET center. Time from injury to admission, time from injury to first debridement, wound closure time, and initial Intra Venous antibiotic time from the injury are an important factor that can be readily amended to improve the infection rate. Whether wound closed before seven days or not were more important factor associated with occurrences of infection.

Keywords: infection, open tibia, fracture, magnitude

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Authors: Arzu Karahan, Loriano Ballarin, Baruch Rinkevich

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Marine invertebrates, the highly diverse phyla of multicellular organisms, represent phenomena that are either not found or highly restricted in the vertebrates. These include phenomena like budding, fission, a fusion of ramets, and high regeneration power, such as the ability to create whole new organisms from either tiny parental fragment, many of which are controlled by totipotent, pluripotent, and multipotent stem cells. Thus, there is very much that can be learned from these organisms on the practical and evolutionary levels, further resembling Darwin's words, “It is not the strongest of the species that survives, nor the most intelligent, but the one most responsive to change”. The ‘stem cell’ notion highlights a cell that has the ability to continuously divide and differentiate into various progenitors and daughter cells. In vertebrates, adult stem cells are rare cells defined as lineage-restricted (multipotent at best) with tissue or organ-specific activities that are located in defined niches and further regulate the machinery of homeostasis, repair, and regeneration. They are usually categorized by their morphology, tissue of origin, plasticity, and potency. The above description not always holds when comparing the vertebrates with marine invertebrates’ stem cells that display wider ranges of plasticity and diversity at the taxonomic and the cellular levels. While marine/aquatic invertebrates stem cells (MISC) have recently raised more scientific interest, the know-how is still behind the attraction they deserve. MISC, not only are highly potent but, in many cases, are abundant (e.g., 1/3 of the entire animal cells), do not locate in permanent niches, participates in delayed-aging and whole-body regeneration phenomena, the knowledge of which can be clinically relevant. Moreover, they have massive hidden potential for the discovery of new bioactive molecules that can be used for human health (antitumor, antimicrobial) and biotechnology. The MARISTEM COST action (Stem Cells of Marine/Aquatic Invertebrates: From Basic Research to Innovative Applications) aims to connect the European fragmented MISC community. Under this scientific umbrella, the action conceptualizes the idea for adult stem cells that do not share many properties with the vertebrates’ stem cells, organizes meetings, summer schools, and workshops, stimulating young researchers, supplying technical and adviser support via short-term scientific studies, making new bridges between the MISC community and biomedical disciplines.

Keywords: aquatic/marine invertebrates, adult stem cell, regeneration, cell cultures, bioactive molecules

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Authors: Soroosh Torabi, Brad Berron, Ren Xu, Christine Trinkle

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Microfluidics integrated with 3D cell culture is a powerful technology to mimic cellular environment, and can be used to study cell activities such as proliferation, migration and response to drugs. This technology has gained more attention in cancer studies over the past years, and many organ-on-a-chip systems have been developed to study cancer cell behaviors in an ex-vivo tumor microenvironment. However, there are still some barriers to adoption which include low throughput, complexity in 3D cell culture integration and limitations on non-optical analysis of cells. In this study, a user-friendly microfluidic multi-well plate was developed to mimic the in vivo tumor microenvironment. The microfluidic platform feeds multiple 3D cell culture sites at the same time which enhances the throughput of the system. The platform uses hydrophobic Cassie-Baxter surfaces created by microchannels to enable convenient loading of hydrogel/cell suspensions into the device, while providing barrier free placement of the hydrogel and cells adjacent to the fluidic path. The microchannels support convective flow and diffusion of nutrients to the cells and a removable lid is used to enable further chemical and physiological analysis on the cells. Different breast cancer cell lines were cultured in the device and then monitored to characterize nutrient delivery to the cells as well as cell invasion and proliferation. In addition, the drug response of breast cancer cell lines cultured in the device was compared to the response in xenograft models to the same drugs to analyze relevance of this platform for use in future drug-response studies.

Keywords: microfluidics, multi-well 3d cell culture, tumor microenvironment, tumor-on-a-chip

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Authors: Nicole Harley, Mahmoud Eid, Abdurahman Tarmal, Vishal Dey

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Histological sampling plays an integral role in the diagnostic process of renal diseases. Percutaneous native renal biopsy is typically performed under ultrasound guidance, with this service usually being provided by nephrologists. In some centers, there is a role for radiologists in performing renal biopsies. Previous comparative studies have demonstrated non-inferiority between outcomes of percutaneous native renal biopsies performed by nephrologists compared with radiologists. We sought to compare biopsy adequacy between nephrologists and radiologists in University Hospital Crosshouse. The online system SERPR (Scottish Electronic Renal Patient Record) contains information pertaining to patients who have undergone renal biopsies. An online search was performed to acquire a list of all patients who underwent renal biopsy between 2013 and 2020 in University Hospital Crosshouse. 355 native renal biopsies were performed in total across this 7-year period. A retrospective analysis was performed on these cases, with records and reports being assessed for: the total number of glomeruli obtained per biopsy, whether the number of glomeruli obtained was adequate for diagnosis, as per an internationally agreed standard, and whether a histological diagnosis was achieved. Nephrologists performed 43.9% of native renal biopsies (n=156) and radiologists performed 56.1% (n=199). The mean number of glomeruli obtained by nephrologists was 17.16+/-10.31. The mean number of glomeruli obtained by radiologists was 18.38+/-10.55. T-test demonstrated no statistically significant difference between specialties comparatively (p-value 0.277). Native renal biopsies are required to obtain at least 8 glomeruli to be diagnostic as per internationally agreed criteria. Nephrologists met these criteria in 88.5% of native renal biopsies (n=138) and radiologists met this criteria in 89.5% (n=178). T-test and Chi-squared analysis demonstrate there was no statistically significant difference between the specialties comparatively (p-value 0.663 and 0.922, respectively). Biopsies performed by nephrologists yielded tissue that was diagnostic in 91.0% (n=142) of sampling. Biopsies performed by radiologists yielded tissue that was diagnostic in 92.4% (n=184) of sampling. T-test and Chi-squared analysis demonstrate there was no statistically significant difference between the specialties comparatively (p-value 0.625 and 0.889, respectively). This project demonstrates that at University Hospital Crosshouse, there is no statistical difference between radiologists and nephrologists in terms of glomeruli acquisition or samples achieving a histological diagnosis. Given the non-inferiority between specialties demonstrated by previous studies and this project, this evidence could support the restructuring of services to allow more renal biopsies to be performed by renal services and allow reallocation of radiology department resources.

Keywords: biopsy, medical imaging, nephrology, radiology

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Authors: S. Buloyan, V. Mamikonyan, H. Hakobyan, H. Harutyunyan, H. Gasparyan

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The liver is the strongest regenerating organ of the organism, and even with 2/3 surgically removed, it can regenerate completely. Hence, liver cirrhosis may only develop when the regenerating system is off. We present the results of a comparative study of structural and functional characteristics of rat liver tissue under the conditions of toxic liver cirrhosis development, induced by carbon tetrachloride, and its prevention/treatment by natural compounds with antioxidant and immune stimulating action. Studies were made on Wister rats, weighing 120~140 g. Grape seeds extracts, separately and in combination with well known anticirrhotic drug ursodeoxycholic acid (ursodiol) have demonstrated effectiveness in prevention of liver cirrhosis development and its treatment.

Keywords: carbon tetrachloride, GSE, liver cirrhosis, prevention, treatment

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Authors: Yu-Chen Hu

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Gene editing by CRISPR and gene regulation by microRNA or CRISPR activation have dramatically changed the way to manipulate cellular gene expression and cell fate. In recent years, various gene editing and gene manipulation technologies have been applied to control stem cell differentiation to enhance tissue regeneration. This research will focus on how to develop CRISPR, CRISPR activation (CRISPRa), CRISPR inhibition (CRISPRi), as well as bi-directional CRISPR-AI gene regulation technologies to control cell differentiation and bone regeneration. Moreover, in this study, CRISPR/Cas13d-mediated RNA editng for miRNA editing and bone regeneration will be discussed.

Keywords: gene therapy, bone regeneration, stem cell, CRISPR, gene regulation

Procedia PDF Downloads 90

Authors: Constantine Uwaremwe, Wenjie Bao, Bachir Goudia Daoura, Sandhya Mishra, Xianxian Zhang, Lingjie Shen, Shangwen Xia, Xiaodong Yang

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Background: Plukenetia volubilis Linneo is an oleaginous plant belonging to the family Euphorbiaceae. Due to its seeds containing a high content of edible oil and rich in vitamins, P. volubilis is cultivated as an economical plant worldwide. However, the cultivation and growth of P. volubilis is challenged by phytopathogen invasion leading to production loss. Methods: In the current study, we tested the pathogenicity of fungal pathogens isolated from root rot infected P. volubilis plant tissues by inoculating them into healthy P. volubilis seedlings. Metagenomic sequencing was used to assess the shift in the fungal community of P. volubilis rhizosphere soil after root rot infection. Results: Four Fusarium isolates and two Rhizopus isolates were found to be root rot causative agents of P. volubilis as they induced typical root rot symptoms in healthy seedlings. The metagenomic sequencing data showed that root rot infection altered the rhizosphere fungal community. In root rot infected soil, the richness and diversity indices increased or decreased depending on pathogens. The four most abundant phyla across all samples were Ascomycota, Glomeromycota, Basidiomycota, and Mortierellomycota. In infected soil, the relative abundance of each phylum increased or decreased depending on the pathogen and functional taxonomic classification. Conclusions: Based on our results, we concluded that Fusarium and Rhizopus species cause root rot infection of P. volubilis. In root rot infected P. volubilis, the shift in the rhizosphere fungal community was pathogen-dependent. These findings may serve as a key point for a future study on the biocontrol of root rot of P. volubilis.

Keywords: fusarium spp., plukenetia volubilis l., rhizopus spp., rhizosphere fungal community, root rot

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Authors: Samantha Meyerholz

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MSKCC offers patients cutting edge cancer care with the highest quality standards. However, many patients and industry members do not realize that the operations of the Immunology Histology Lab (IHC) are the backbone for carrying out this mission. The IHC lab manufactures blocks and slides containing critical tissue samples that will be read by a Pathologist to diagnose and dictate a patient’s treatment course. The lab processes 200 requests daily, leading to the generation of approximately 2,000 slides and 1,100 blocks each day. Lab material is transported through labeling, cutting, staining and sorting manufacturing stations, while being managed by multiple techs throughout the space. The quality of the stain as well as wait times associated with processing requests, is directly associated with patients receiving rapid treatments and having a wider range of care options. This project aims to improve slide request turnaround time for rush and non-rush cases, while increasing the quality of each request filled (no missing slides or poorly stained items). Rush cases are to be filled in less than 24 hours, while standard cases are allotted a 48 hour time period. Reducing turnaround times enable patients to communicate sooner with their clinical team regarding their diagnosis, ultimately leading faster treatments and potentially better outcomes. Additional project goals included streamlining tech and material workflow, while reducing waste and increasing efficiency. This project followed a DMAIC structure with emphasis on lean and ergonomic principles that could be integrated into an evolving lab culture. Load times and batching processes were analyzed using process mapping, FMEA analysis, waste analysis, engineering observation, 5S and spaghetti diagramming. Reduction of lab technician movement as well as their body position at each workstation was of top concern to pathology leadership. With new equipment being brought into the lab to carry out workflow improvements, screen and tool placement was discussed with the techs in focus groups, to reduce variation and increase comfort throughout the workspace. 5S analysis was completed in two phases in the IHC lab, helping to drive solutions that reduced rework and tech motion. The IHC lab plans to continue utilizing these techniques to further reduce the time gap between tissue analysis and cancer care.

Keywords: engineering, ergonomics, healthcare, lean

Procedia PDF Downloads 223

Authors: Fahimeh Palizban, Farshad Noravesh, Amir Hossein Saeidian, Mahya Mehrmohamadi

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In the recent decade, the emergence of liquid biopsy has significantly improved cancer monitoring and detection. Dying cells, including those originating from tumors, shed their DNA into the blood and contribute to a pool of circulating fragments called cell-free DNA. Accordingly, identifying the tissue origin of these DNA fragments from the plasma can result in more accurate and fast disease diagnosis and precise treatment protocols. Open chromatin regions are important epigenetic features of DNA that reflect cell types of origin. Profiling these features by DNase-seq, ATAC-seq, and histone ChIP-seq provides insights into tissue-specific and disease-specific regulatory mechanisms. There have been several studies in the area of cancer liquid biopsy that integrate distinct genomic and epigenomic features for early cancer detection along with tissue of origin detection. However, multimodal analysis requires several types of experiments to cover the genomic and epigenomic aspects of a single sample, which will lead to a huge amount of cost and time. To overcome these limitations, the idea of predicting OCRs from WGS is of particular importance. In this regard, we proposed a computational approach to target the prediction of open chromatin regions as an important epigenetic feature from cell-free DNA whole genome sequence data. To fulfill this objective, local sequencing depth will be fed to our proposed algorithm and the prediction of the most probable open chromatin regions from whole genome sequencing data can be carried out. Our method integrates the signal processing method with sequencing depth data and includes count normalization, Discrete Fourie Transform conversion, graph construction, graph cut optimization by linear programming, and clustering. To validate the proposed method, we compared the output of the clustering (open chromatin region+, open chromatin region-) with previously validated open chromatin regions related to human blood samples of the ATAC-DB database. The percentage of overlap between predicted open chromatin regions and the experimentally validated regions obtained by ATAC-seq in ATAC-DB is greater than 67%, which indicates meaningful prediction. As it is evident, OCRs are mostly located in the transcription start sites (TSS) of the genes. In this regard, we compared the concordance between the predicted OCRs and the human genes TSS regions obtained from refTSS and it showed proper accordance around 52.04% and ~78% with all and the housekeeping genes, respectively. Accurately detecting open chromatin regions from plasma cell-free DNA-seq data is a very challenging computational problem due to the existence of several confounding factors, such as technical and biological variations. Although this approach is in its infancy, there has already been an attempt to apply it, which leads to a tool named OCRDetector with some restrictions like the need for highly depth cfDNA WGS data, prior information about OCRs distribution, and considering multiple features. However, we implemented a graph signal clustering based on a single depth feature in an unsupervised learning manner that resulted in faster performance and decent accuracy. Overall, we tried to investigate the epigenomic pattern of a cell-free DNA sample from a new computational perspective that can be used along with other tools to investigate genetic and epigenetic aspects of a single whole genome sequencing data for efficient liquid biopsy-related analysis.

Keywords: open chromatin regions, cancer, cell-free DNA, epigenomics, graph signal processing, correlation clustering

Procedia PDF Downloads 150

Authors: Keivan Jamshidi, Fateme Behbahani, Sara Omidi, Nadia Shahi, Alireza Farkhonde

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Amyloidosis represents a heterogenous group of diseases that have in common the deposition of fibrils composed of proteins of beta-pleated sheet structure, which can be specifically identified by histochemistry using the Congo red or similar stains. Between October 2013 to April 2014 (6 months) different patterns of renal amyloidosis was diagnosed on histopathological examination of kidneys belong to 196 out of 7065 slaughtered sheep subjected to postmortem examination. Microscopic examination of renal tissue sections stained with H&E and CR staining techniques revealed 3 patterns of renal amyloid deposition; including glomerular (22.72%), medullary (68.18%), and vascular (9.09%) were recognized. Renal medullary amyloidosis (RMA) was detected as the most prevalence pattern of renal amyloidosis in domestic sheep.

Keywords: sheep, amyloidosis, kidney, slaughterhouse

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Authors: H. Al-Khalifa, A. Al-Nasser

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The main immune organs in poultry are the thymus, spleen and bursa of Fabricius. During an immune response, mature lymphocytes and other immune cells interact with antigens in these tissues. Consequently, the mass of these organs can in some cases indicate immune status. The objective of the current study was to investigate the effect of feeding flaxseed on immune tissue weights. Cobb 500 broiler chickens were fed flaxseed at 15%, the control diet did not contain any flaxseed. Results showed that dietary supplementation with flaxseed did not affect the weights of the spleens of broiler chickens. However, it significantly lowered bursa weights (p<0.01), compared to the control diet. In addition, the bursae were thinner in appearance compared with bursii from chickens fed the control diets.

Keywords: bursa of fabricius, flaxseed, spleen, thymus

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Authors: M. Y. Lee, D. H. Shin, S. H. Park, W.C. Ham, S.K. Ko, C. G. Song

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Photoacoustic Tomography (PAT) is a promising medical imaging modality that combines optical imaging contrast with the spatial resolution of ultrasound imaging. It can also distinguish the changes in biological features. But, real-time PAT system should be confirmed due to photoacoustic effect for tissue. Thus, we have developed a real-time PAT system using a custom-developed data acquisition board and ultrasound linear probe. To evaluate performance of our system, phantom test was performed. As a result of those experiments, the system showed satisfactory performance and its usefulness has been confirmed. We monitored the degradation of inflammation which induced on the rat’s kidney using real-time PAT.

Keywords: photoacoustic tomography, inflammation detection, rat, kidney, contrast agent, ultrasound

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Authors: Dipali Dhawan

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Cancerous epithelial cells are confined to a primary site by the continued expression of adhesion molecules and the intact basal lamina. However, as the cancer progresses some cells are believed to undergo an epithelial-mesenchymal transition (EMT) event, leading to increased motility, invasion and, ultimately, metastasis of the cells from the primary tumour to secondary sites within the body. These disseminated cancer cells need the ability to self-renew, as stem cells do, in order to establish and maintain a heterogeneous metastatic tumour mass. Identification of the specific subpopulation of cancer stem cells amenable to the process of metastasis is highly desirable. In this study, we have isolated and characterized cancer stem cells from luminal and basal breast cancer cell lines (MDA-MB-231, MDA-MB-453, MDA-MB-468, MCF7 and T47D) on the basis of cell surface markers CD44 and CD24; as well as Side Populations (SP) using Hoechst 33342 dye efflux. The isolated populations were analysed for epithelial and mesenchymal markers like E-cadherin, N-cadherin, Sfrp1 and Vimentin by Western blotting and Immunocytochemistry. MDA-MB-231 cell lines contain a major population of CD44+CD24- cells whereas MCF7, T47D and MDA-MB-231 cell lines show a side population. We observed higher expression of N-cadherin in MCF-7 SP cells as compared to MCF-7NSP (Non-side population) cells suggesting that the SP cells are mesenchymal like cells and hence express increased N-cadherin with stem cell-like properties. There was an expression of Sfrp1 in the MCF7- NSP cells as compared to no expression in MCF7-SP cells, which suggests that the Wnt pathway is expressed in the MCF7-SP cells. The mesenchymal marker Vimentin was expressed only in MDA-MB-231 cells. Hence, understanding the breast cancer heterogeneity would enable a better understanding of the disease progression and therapeutic targeting.

Keywords: cancer stem cells, epithelial to mesenchymal transition, biomarkers, breast cancer

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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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Authors: Devinder Kaur Sugga, Ekamdeep Kaur, Jaspreet Kaur, C. Rajesh, Preeti Rajesh, Harsimran Kaur

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Withanolides are steroidal lactones and are highly oxygenated phytoconstituents that can be developed as potential anti-carcinogenic agents. The two main withanolides, namely Withaferin A and Withanolides D, have been extensively studied for their pharmacological activities. Both these withanolides are present in the Withania somnifera (WS) leaves belonging to the family Solanaceae, also known as “Indian ginseng .”In this study effects of WS leaf extract on the MCF7 breast cancer cell line were investigated by performing a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to evaluate the cytotoxic effects and in vitro wound-healing assay to study the effect on cancer cell migration. Our data suggest WS extracts have cytotoxic effects and are effective anti-migrating agents and thus can be a source of potential candidates for the development of potential agents against metastasis. Thus, it can be a source of potential candidates for the development of potential agents against metastasis. Insight into these results, the in-silico approach to identify the possible protein targets interacting with withanolides was taken. Protein kinase C alpha (PKCα) was among the selected 5 top-ranked target proteins identified by the Swiss Target Prediction tool. PKCα is known to promote the growth and invasion of cancer cells and is being evaluated as a prognostic biomarker and therapeutic target in clinically aggressive tumors. Molecular docking of Withaferin A and Withanolides D was performed using AutoDock Vina. Both the bioactive compounds interacted with PKCα. The targets predicted using this approach will serve as leads for the possible therapeutic potential of withanolides, the bioactive ingredients of WS extracts, as anti-cancer drugs.

Keywords: withania somnifera, withaferin A, withanolides D, PKCα

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Authors: K. J. Keerthi, Vasundhara Kamineni, A. Ravi Shanker, T. Rammurthy, A. Vijaya Lakshmi, Q. Hasan

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Pancreatic β-cells are the predominant insulin-producing cell types within the Islets of Langerhans and insulin is the primary hormone which regulates carbohydrate and fat metabolism. Apoptosis of β-cells or insufficient insulin production leads to Diabetes Mellitus (DM). Current therapy for diabetes includes either medical management or insulin replacement and regular monitoring. Replacement of β- cells is an attractive treatment option for both Type-1 and Type-2 DM in view of the recent paper which indicates that β-cells apoptosis is the common underlying cause for both the Types of DM. With the development of Edmonton protocol, pancreatic β-cells allo-transplantation became possible, but this is still not considered as standard of care due to subsequent requirement of lifelong immunosuppression and the scarcity of suitable healthy organs to retrieve pancreatic β-cell. Fetal pancreatic cells from abortuses were developed as a possible therapeutic option for Diabetes, however, this posed several ethical issues. Hence, in the present study Mesenchymal stem cells (MSCs) were differentiated into insulin producing cells which were isolated from Human Umbilical cord (HUC) tissue. MSCs have already made their mark in the growing field of regenerative medicine, and their therapeutic worth has already been validated for a number of conditions. HUC samples were collected with prior informed consent as approved by the Institutional ethical committee. HUC (n=26) were processed using a combination of both mechanical and enzymatic (collagenase-II, 100 U/ml, Gibco ) methods to obtain MSCs which were cultured in-vitro in L-DMEM (Low glucose Dulbecco's Modified Eagle's Medium, Sigma, 4.5 mM glucose/L), 10% FBS in 5% CO2 incubator at 37°C. After reaching 80-90% confluency, MSCs were characterized with Flowcytometry and Immunocytochemistry for specific cell surface antigens. Cells expressed CD90+, CD73+, CD105+, CD34-, CD45-, HLA-DR-/Low and Vimentin+. These cells were differentiated to β-cells by using H-DMEM (High glucose Dulbecco's Modified Eagle's Medium,25 mM glucose/L, Gibco), β-Mercaptoethanol (0.1mM, Hi-Media), basic Fibroblast growth factor (10 µg /L,Gibco), and Nicotinamide (10 mmol/L, Hi-Media). Pancreatic β-cells were confirmed by positive Dithizone staining and were found to be functionally active as they released 8 IU/ml insulin on glucose stimulation. Isolating MSCs from usually discarded, abundantly available HUC tissue, expanding and differentiating to β-cells may be the most feasible cell therapy option for the millions of people suffering from DM globally.

Keywords: diabetes mellitus, human umbilical cord, mesenchymal stem cells, differentiation

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Authors: Ferreira R., Rocha K.

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Ozone therapy is indicated for improving skin aesthetics, adjusting oxidative tissue levels, increasing collagen production, and even skin volumizing. This paper aims to carry out a case report that demonstrates the positive results of ozone therapy in association with superficial peeling. The application in association showed positive results for bio-stimulating activities in the reported case demonstrating to be a viable clinical technique. The bio-stimulating effect of ozone therapy in association with peeling is a promising aesthetic therapeutic modality with fast and safe results as an aesthetic therapeutic option.

Keywords: bio-stimulating effect, ozone therapy, neocollagenesis, peeling

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Authors: Lubna M. Al-Otaibi

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Mucocele is a common benign neoplasm of the oral cavity and the most common after fibroma. The lesion develops as a result of retention or extravasation of mucous material from minor salivary glands. Extravasation type of mucocele results from trauma and mostly occurs in the lower lip of young patients. The various treatment options available for the treatment of mucocele are associated with a relatively high incidence of recurrence making surgical intervention necessary for a permanent cure. The conventional surgical procedure, however, arouses apprehension in the patient and is associated with bleeding and postoperative pain. Recently, treatment of mucocele with lasers has become a viable treatment option. Various types of lasers are being used and are preferable over the conventional surgical procedure as they provide good hemostasis, reduced postoperative swelling and pain, reduced bacterial population, lesser need for suturing, faster healing and low recurrence rates. Er,Cr:YSGG is a solid-state laser with great affinity to water molecule. Its hydrokinetic cutting action allows it to work effectively on hydrated tissues without any thermal damage. However, up to date, only a few studies have reported its use in the removal of lip mucocele, especially in children. In this case, a 6 year old female patient with history of trauma to the lower lip presented with a soft, sessile, whitish-bluish 4 mm papule. The lesion was present for approximately four months and was fluctuant in size. The child developed a habit of biting the lesion causing injury, bleeding and discomfort. Surgical excision under local anaesthesia was performed using 2780 nm Er,Cr:YSGG Laser (WaterLase iPlus, Irvine, CA) with a Gold handpiece and MZ6 tip (3.5w, 50 Hz, 20% H2O, 20% Air, S mode). The tip was first applied in contact mode with focused beam using the Circumferential Incision Technique (CIT) to excise the tissue followed by the removal of the underlying causative minor salivary gland. Bleeding was stopped using Laser Dry Bandage setting (0.5w, 50 Hz, 1% H2O, 20% Air, S mode) and no suturing was needed. Safety goggles were worn and high-speed suction was used for smoke evacuation. Mucocele excision using 2780 nm Er,Cr:YSGG laser was rapid, easy to perform with excellent precision and allowed for histopathological examination of the excised tissue. The patient was comfortable and there were minimum bleeding and no sutures, postoperative pain, scarring or recurrence. Laser assisted mucocele excision appears to have efficient and reliable benefits in young patients and should be considered as an alternative to conventional surgical and non-surgical techniques.

Keywords: Erbium, excision, laser, lip, mucocele

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Authors: Meli̇sa Aşci, Berk Kiliç, Emine Ulusoy

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Ingestion of microplastics in humans has been increasing rapidly, as such hazardous materials are abundant in multiple food products, specifically milks. With increasing consumption rates, humans have been ingesting microplastics on a daily basis, making them prone to be intoxicated and even cause the disruption of intracellular pathways and liver cell disruption, and eventually tissue and organ damage. In this experiment, different milk types(animal-based and plant-based) were tested for microplastics. Results showed that animal-based milks contained a higher concentration of microplastics compared to plant-based milks. Research has shown that in addition to causing health issues in humans, microplastics can also affect livestock animals and plants.

Keywords: microplastics, plant-based milks, animal-based milks, preventive nutrition

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Authors: Chang Liang, Weizhi Gong, Yan Zhang

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Purpose: Hepatocellular carcinoma (HCC) is a malignant tumor with a high mortality rate and poor prognosis worldwide. Murine double minute 2 (MDM2) regulates the tumor suppressor p53, increasing cancer risk and accelerating tumor progression. Speckle-type POX virus and zinc finger protein (SPOP), a key of subunit of Cullin-Ring E3 ligase, inhibits tumor genesis and progression by the ubiquitination of its downstream substrates. This study aimed to clarify whether SPOP and MDM2 are mutually regulated in HCC and the correlation between SPOP and MDM2 and the prognosis of HCC patients. Methods: First, the expression of SPOP and MDM2 in HCC tissues were detected by TCGA database. Then, 53 paired samples of HCC tumor and adjacent tissues were collected to evaluate the expression of SPOP and MDM2 using immunohistochemistry. Chi-square test or Fisher’s exact test were used to analyze the relationship between clinicopathological features and the expression levels of SPOP and MDM2. In addition, Kaplan‒Meier curve analysis and log-rank test were used to investigate the effects of SPOP and MDM2 on the survival of HCC patients. Last, the Multivariate Cox proportional risk regression model analyzed whether the different expression levels of SPOP and MDM2 were independent risk factors for the prognosis of HCC patients. Results: Bioinformatics analysis revealed the low expression of SPOP and high expression of MDM2 were related to worse prognosis of HCC patients. The relationship between the expression of SPOP and MDM2 and tumor stem-like features showed an opposite trend. The immunohistochemistry showed the expression of SPOP protein was significantly downregulated while MDM2 protein significantly upregulated in HCC tissue compared to that in para-cancerous tissue. Tumors with low SPOP expression were related to worse T stage and Barcelona Clinic Liver Cancer (BCLC) stage, but tumors with high MDM2 expression were related to worse T stage, M stage, and BCLC stage. Kaplan–Meier curves showed HCC patients with high SPOP expression and low MDM2 expression had better survival than those with low SPOP expression and high MDM2 expression (P < 0.05). A multivariate Cox proportional risk regression model confirmed that a high MDM2 expression level was an independent risk factor for poor prognosis in HCC patients (P <0.05). Conclusion: The expression of SPOP protein was significantly downregulated, while the expression of MDM2 significantly upregulated in HCC. The low expression of SPOP and high expression. of MDM2 were associated with malignant progression and poor prognosis of HCC patients, indicating a potential therapeutic target for HCC patients.

Keywords: hepatocellular carcinoma, murine double minute 2, speckle-type POX virus and zinc finger protein, ubiquitination

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Authors: Yuchen LI, Martyn Kurr, Peter Golyshin

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Biological invasion is a global problem. Invasive species can threaten local ecosystems by competing for resources, consuming local species, and reproducing faster than natives. Sargassum muticum is an invasive algae in the UK. It negatively impacts local algae through overshading and can cause reductions in local biodiversity. One possibility for its success is herbivore release. According to the Enemy Release Hypothesis, invasives are less impacted by local herbivores than natives. In many species, gastrointestinal (GI) tract microbes have been found as a key factor in food preference and similar mechanisms may exist in the relationship between local consumers and S. muticum. Some populations of native Littorina snails accept S. muticum as a food source, while others avoid it. This project aims to establish the relationship between GI tract microbes and the feeding preferences of L. littorea, when offered both native algae and S. muticum. Individuals of L. littorea from a site invaded by S. muticum around 18 years ago were compared to those from an un-invaded site nearby. Sargassum-experienced snails are more likely to consume it than those naïve, and pronounced differences were found in the GI-tract microbial communities through 16S (prokaryote) and 18S (eukaryote) sequencing. Sargassum-naïve snails were then exposed to a faecal pellets from experienced snails to ‘inoculate’ them with microbes from the exposed snails. Preliminary results suggest these faecal-pellet-exposed but otherwise Sargassum-naïve snails subsequently begun consuming S. muticum. It is unclear if these results are due to genuine changes in GI-tract microbes or through some other mechanism, such as behavioural responses to chemical cues in the faecal pellets, but these results are nevertheless of significance for invasive ecology, suggesting that foraging preferences for an invasive prey type are malleable and possibly programmable in laboratory settings.

Keywords: invasive algae, sea snails, gut microbiome, biocontrol

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Authors: Gleb Fomin, Kairat Tabynov, Nurkeldy Turebekov, Dinara Turegeldiyeva, Rinat Islamov

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The level of major cytokines in the blood of patients with COVID-19 varies greatly depending on age, gender, duration and severity of infection, and comorbidity. There are two clinically significant cytokines, IL-6 and TNF-α, which increase in levels in patients with severe COVID-19. However, in a model of COVID-19 in hamsters, TNF-α levels are unchanged or reduced, while the expression of other cytokines reflects the profile of cytokines found in patients’ plasma. The aim of our study was to evaluate the relationship between the level of cytokines in the blood, lungs, and lung damage in the model of the Syrian hamster (Mesocricetus auratus) infected with the SARS-CoV-2 strain. The study used outbred female and male Syrian hamsters (n=36, 4 groups) weighing 80-110 g and 5 months old (protocol IACUC, #4, 09/22/2020). Animals were infected intranasally with the hCoV-19/Kazakhstan/KazNAU-NSCEDI-481/2020 strain and euthanized at 3 d.p.i. The level of cytokines IL-6, TNF-α, IFN-α, and IFN-γ was determined by ELISA MyBioSourse (USA) for hamsters. Lung samples were subjected to histological processing. The presence of pathological changes in histological preparations was assessed on a 3-point scale. The work was carried out in the ABSL-3 laboratory. The data were analyzed in GraphPad Prism 6.00 (GraphPad Software, La Jolla, California, USA). The work was supported by the MES RK grant (AP09259865). In the blood, the level of TNF-α increased in males (p=0.0012) and IFN-γ in males and females (p=0.0001). On the contrary, IFN-α production decreased (p=0.0006). Only TNF-α level increased in lung tissues (p=0.0011). Correlation analysis showed a negative relationship between the level of IL-6 in the blood and lung damage in males (r -0.71, p=0.0001) and females (r-0.57, p=0.025). On the contrary, in males, the level of IL-6 in the lungs and score is positively correlated (r 0.80, p=0.01). The level of IFN-γ in the blood (r -0.64, p=0.035) and lungs (r-0.72, p=0.017) in males has a negative correlation with lung damage. No links were found for TNF-α and IFN-α. The study showed a positive association between lung injury and tissue levels of IL-6 in male hamsters. It is known that in humans, high concentrations of IL-6 in the lungs are associated with suppression of cellular immunity and, as a result, with an increase in the severity of COVID-19. TNF-α and IFN-γ play a key role in the pathogenesis of COVID-19 in hamsters. However, the mechanisms of their activity require more detailed study. IFN-α plays a lesser role in direct lung injury in a Syrian hamster model. We have shown the significance of tissue IL-6 and IFN-γ as predictors of the severity of lung damage in COVID-19 in the Syrian hamster model. Changes in the level of cytokines in the blood may not always reflect pathological processes in the lungs with COVID-19.

Keywords: syrian hamster, COVID-19, cytokines, biological model

Procedia PDF Downloads 92

Authors: Alina Murtishcheva

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The tendency of globalization, challenges to democracy and peace caused by the Russian invasion of Ukraine, and other global conflicts require searching general orientations of governmental development, including local government. The formation of a common theoretical framework for local government guarantees not only of harmonisation of European legislation but also creates prerequisites for the integration of new members into the European Union. One of the most important milestones of such a theoretical framework is the concept of decentralization. Decentralization as a phenomenon is characteristic of most European Union countries at different historical stages. For Ukraine, as a country that has clearly defined a European integration vector of development, understanding not only the legal but also the theoretical basis of decentralisation processes in European countries is an important prerequisite for further reforms. Decentralisation takes different forms, which leads to a variety of understandings in doctrine and, consequently, different interpretations in national legislation. Despite of this, decentralisation is based on common ideas and values such as democracy, participation, the rule of law, and proximity government that are shared by all EU member states. Nevertheless, not all EU countries are currently implementing broad decentralization in their political and legal practices. Some countries are gradually moving in this direction, while others remain quite centralised. There is also a new, insufficiently studied trend today – recentralisation, which can be broadly defined as the strengthening of centralization tendencies in countries that were considered to be decentralized. Consequently, an exploratory theoretical study is needed to identify how the concept of decentralization is combined with the recentralization tendency in EU member states. The purpose of this study is to empirically analyse scientific approaches to the concept of “decentralisation”, to highlight the tendency of recentralisation and its consequences, to analyse Ukraine's experience in the field of decentralisation of public power, and to outline the prospects for further development of Ukrainian legislation in this area.

Keywords: centralization, decentralization, local government, recentralization, reforms

Procedia PDF Downloads 75

Authors: Sara Vora (Hoxha)

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Defamation, invasion of privacy, and cyberbullying have all increased in tandem with the growth of the internet. European international private law may struggle to deal with such transgressions if they occur in many jurisdictions. The current study examines how effectively the legal system of European international private law addresses abuses of personality rights in cyberspace. The study starts by discussing how established legal frameworks are being threatened by online personality rights abuses. The article then looks into the rules and regulations of European international private law that are in place to handle overseas lawsuits. This article examines the different elements that courts evaluate when deciding which law to use in a particular case, focusing on the concepts of jurisdiction, choice of law, and recognition and execution of foreign judgements. Next, the research analyses the function of the European Union in preventing and punishing online personality rights abuses. Key pieces of law that control the collecting and processing of personal data on the Internet, including the General Data Protection Regulation (GDPR) and the e-Commerce Directive, are discussed. In addition, this article investigates how the ECtHR handles cases involving the infringement of personal freedoms, including privacy and speech. The article finishes with an assessment of how well the legal framework of European international private law protects individuals' right to privacy online. It draws attention to problems with the present legal structure, such as the inability to enforce international judgements, the inconsistency between national laws, and the necessity for stronger measures to safeguard people' rights online. This paper concludes that while European international private law provides a useful framework for dealing with violations of personality rights online, further harmonisation and stronger enforcement mechanisms are necessary to effectively protect individuals' rights in the digital age.

Keywords: European international private law, personality rights, internet, jurisdiction, cross-border disputes, data protection

Procedia PDF Downloads 75

Authors: E. Nakamachi, K. Matsumoto, K. Yamamoto, Y. Morita, H. Sakamoto

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In recently, electromagnetic and mechanical stimulations have been recognized as the effective extracellular environment stimulation technique to enhance the defected peripheral nerve tissue regeneration. In this study, we developed a new hybrid bioreactor by adopting 50 Hz uniform alternative current (AC) magnetic stimulation and 4% strain mechanical stimulation. The guide tube for nerve regeneration is mesh structured tube made of biodegradable polymer, such as polylatic acid (PLA). However, when neural damage is large, there is a possibility that peripheral nerve undergoes necrosis. So it is quite important to accelerate the nerve tissue regeneration by achieving enhancement of nerve axonal extension rate. Therefore, we try to design and fabricate the system that can simultaneously load the uniform AC magnetic field stimulation and the stretch stimulation to cells for enhancement of nerve axonal extension. Next, we evaluated systems performance and the effectiveness of each stimulation for rat adrenal pheochromocytoma cells (PC12). First, we designed and fabricated the uniform AC magnetic field system and the stretch stimulation system. For the AC magnetic stimulation system, we focused on the use of pole piece structure to carry out in-situ microscopic observation. We designed an optimum pole piece structure using the magnetic field finite element analyses and the response surface methodology. We fabricated the uniform AC magnetic field stimulation system as a bio-reactor by adopting analytically determined design specifications. We measured magnetic flux density that is generated by the uniform AC magnetic field stimulation system. We confirmed that measurement values show good agreement with analytical results, where the uniform magnetic field was observed. Second, we fabricated the cyclic stretch stimulation device under the conditions of particular strains, where the chamber was made of polyoxymethylene (POM). We measured strains in the PC12 cell culture region to confirm the uniform strain. We found slightly different values from the target strain. Finally, we concluded that these differences were allowable in this mechanical stimulation system. We evaluated the effectiveness of each stimulation to enhance the nerve axonal extension using PC12. We confirmed that the average axonal extension length of PC12 under the uniform AC magnetic stimulation was increased by 16 % at 96 h in our bio-reactor. We could not confirm that the axonal extension enhancement under the stretch stimulation condition, where we found the exfoliating of cells. Further, the hybrid stimulation enhanced the axonal extension. Because the magnetic stimulation inhibits the exfoliating of cells. Finally, we concluded that the enhancement of PC12 axonal extension is due to the magnetic stimulation rather than the mechanical stimulation. Finally, we confirmed that the effectiveness of the uniform AC magnetic field stimulation for the nerve axonal extension using PC12 cells.

Keywords: nerve cell PC12, axonal extension, nerve regeneration, electromagnetic-mechanical stimulation, bioreactor

Procedia PDF Downloads 265

Authors: Zeynep Busra Velioglu, Deniz Pulat, Beril Demirbakan, Burak Ozcan, Ece Bayrak, Cevat Erisken

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Bone tissue has the ability to perform a wide array of functions including providing posture, load-bearing capacity, protection for the internal organs, initiating hematopoiesis, and maintaining the homeostasis of key electrolytes via calcium/phosphate ion storage. The most common cause for bone defects is extensive trauma and subsequent infection. Bone tissue has the self-healing capability without a scar tissue formation for the majority of the injuries. However, some may result with delayed union or fracture non-union. Such cases include reconstruction of large bone defects or cases of compromised regenerative process as a result of avascular necrosis and osteoporosis. Several surgical methods exist to treat bone defects, including Ilizarov method, Masquelete technique, growth factor stimulation, and bone replacement. Unfortunately, these are technically demanding and come with noteworthy disadvantages such as lengthy treatment duration, adverse effects on the patient’s psychology, repeated surgical procedures, and often long hospitalization times. These limitations associated with surgical techniques make bone substitutes an attractive alternative. Here, it was hypothesized that a 3D printed scaffold will mimic trabecular bone in terms of biomechanical properties and that such scaffolds will support cell attachment and survival. To test this hypothesis, this study aimed at fabricating poly(lactic acid), PLA, structures using 3D printing technology for trabecular bone defects, characterizing the scaffolds and comparing with bovine trabecular bone. Capacity of scaffolds on human bone marrow stem cell (hBMSC) attachment and survival was also evaluated. Cubes with a volume of 1 cm³ having pore sizes of 0.50, 1.00 and 1.25 mm were printed. The scaffolds/grafts were characterized in terms of porosity, contact angle, compressive mechanical properties as well cell response. Porosities of the 3D printed scaffolds were calculated based on apparent densities. For contact angles, 50 µl distilled water was dropped over the surface of scaffolds, and contact angles were measured using ‘Image J’ software. Mechanical characterization under compression was performed on scaffolds and native trabecular bone (bovine, 15 months) specimens using a universal testing machine at a rate of 0.5mm/min. hBMSCs were seeded onto the 3D printed scaffolds. After 3 days of incubation with fully supplemented Dulbecco’s modified Eagle’s medium, the cells were fixed using 2% formaldehyde and glutaraldehyde mixture. The specimens were then imaged under scanning electron microscopy. Cell proliferation was determined by using EZQuant dsDNA Quantitation kit. Fluorescence was measured using microplate reader Spectramax M2 at the excitation and emission wavelengths of 485nm and 535nm, respectively. Findings suggested that porosity of scaffolds with pore dimensions of 0.5mm, 1.0mm and 1.25mm were not affected by pore size, while contact angle and compressive modulus decreased with increasing pore size. Biomechanical characterization of trabecular bone yielded higher modulus values as compared to scaffolds with all pore sizes studied. Cells attached and survived in all surfaces, demonstrating higher proliferation on scaffolds with 1.25mm pores as compared with those of 1mm. Collectively, given lower mechanical properties of scaffolds as compared to native bone, and biocompatibility of the scaffolds, the 3D printed PLA scaffolds of this study appear as candidate substitutes for bone repair and regeneration.

Keywords: 3D printing, biomechanics, bone repair, stem cell

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Authors: Sara R. Knigge, Sugat R. Tuladhar, Hans-Klaus HöFfler, Tobias Schilling, Tim Kaufeld, Axel Haverich

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Tissue engineered (TE) heart valves based on degradable electrospun fiber scaffold represent a promising approach to overcome the known limitations of mechanical or biological prostheses. But the mechanical stress in the high-pressure system of the human circulation is a severe challenge for the delicate materials. Hence, the prediction of the scaffolds` in vivo degradation kinetics must be as accurate as possible to prevent fatal events in future animal or even clinical trials. Therefore, this study investigates whether long-term testing in full blood provides more meaningful results regarding the degradation behavior than conventional tests in simulated body fluids (SBF) or Phosphate Buffered Saline (PBS). Fiber mats were produced from a polycaprolactone (PCL)/tetrafluoroethylene solution by electrospinning. The morphology of the fiber mats was characterized via scanning electron microscopy (SEM). A maximum physiological degradation environment utilizing a test set-up with porcine full blood was established. The set-up consists of a reaction vessel, an oxygenator unit, and a roller pump. The blood parameters (pO2, pCO2, temperature, and pH) were monitored with an online test system. All tests were also carried out in the test circuit with SBF and PBS to compare conventional degradation media with the novel full blood setting. The polymer's degradation is quantified by SEM picture analysis, differential scanning calorimetry (DSC), and Raman spectroscopy. Tensile and cyclic loading tests were performed to evaluate the mechanical integrity of the scaffold. Preliminary results indicate that PCL degraded slower in full blood than in SBF and PBS. The uptake of water is more pronounced in the full blood group. Also, PCL preserved its mechanical integrity longer when degraded in full blood. Protein absorption increased during the degradation process. Red blood cells, platelets, and their aggregates adhered on the PCL. Presumably, the degradation led to a more hydrophilic polymeric surface which promoted the protein adsorption and the blood cell adhesion. Testing degradable implants in full blood allows for developing more reliable scaffold materials in the future. Material tests in small and large animal trials thereby can be focused on testing candidates that have proven to function well in an in-vivo-like setting.

Keywords: Electrospun scaffold, full blood degradation test, long-term polymer degradation, tissue engineered aortic heart valve

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Authors: Logan Carmichael

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As the world’s governments seek to increasingly digitize their service provisions, there exists a subsequent and fully valid concern about the security underpinning these digital governance provisions. Estonia, a small and innovative Baltic nation, has been refining both its digital governance structure and cybersecurity mechanisms for over three decades and has been praised as global ‘best practice’ in both fields. However, the security of the Estonian digital governance system has been ever-evolving and significantly shaped by cybersecurity crises. This paper examines said crises – 2007 cyberattacks on Estonian government, banks, and news media; the 2017 e-ID crisis; the ongoing COVID-19 pandemic; and the 2022 Russian invasion of Ukraine – and how governance decision-making following these crises has shaped the cybersecurity of the digital governance structure in Estonia. This paper employs a blended constructivist and historical institutionalist theoretical approach as a useful means to view governance and decision-making in the wake of cybersecurity incidents affecting the Estonian digital governance structure. Together, these theoretical groundings frame the topics of cybersecurity and digital governance in an Estonian context through a lens of ideation and experience, as well as institutional path dependencies over time and cybersecurity crises as critical junctures to study. Furthermore, this paper takes a qualitative approach, employing discourse analysis, policy analysis, and elite interviewing of Estonian officials involved in digital governance and cybersecurity in order to glean nuanced perspectives into the processes that followed these four crises. Ultimately, the results of this paper will offer insight into how governments undertake policy-driven change following cybersecurity crises to ensure sufficient security of their digitized service provisions. This paper’s findings are informative not only in continued decision-making in the Estonian system but also in other states currently implementing a digital governance structure, for which security mechanisms are of the utmost importance.

Keywords: cybersecurity, digital governance, Estonia, crisis management, governance in crisis

Procedia PDF Downloads 112