Search results for: cellular and castellated beams

Authors: Hema V. Vanar, R. K. Soni, N. D. Shah

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Slabs supported directly on columns without beams are known as Flat slabs. Flat slabs are highly versatile elements widely used in construction, providing minimum depth, fast construction and allowing flexible column grids. The main objective of this thesis is comparison of analytical method and soft ware for analysis of flat slab subjected to various parametric loadings. Study presents analysis of flat slab is performed under different types of gravity.

Keywords: fat slab, parametric load, analysis, software

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Authors: Anjana R. Menon, Anjana Bhasi

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Development of highways and railways play crucial role in a nation’s economic growth. While rigid concrete pavements are durable with high load bearing characteristics, growing economies mostly rely on flexible pavements which are easier in construction and more economical. The strength of flexible pavement is based on the strength of subgrade and load distribution characteristics of intermediate granular layers. In this scenario, to simultaneously meet economy and strength criteria, it is imperative to strengthen and stabilize the load transferring layers, namely subbase and base. Geosynthetic reinforcement in planar and cellular forms have been proven effective in improving soil stiffness and providing a stable load transfer platform. Studies have proven the relative superiority of cellular form-geocells over planar geosynthetic forms like geogrid, owing to the additional confinement of infill material and pocket effect arising from vertical deformation. Hence, the present study investigates the efficiency of geocells over single/multiple layer geogrid reinforcements by a series of three-dimensional model analyses of a flexible pavement section under a standard repetitive wheel load. The stress transfer mechanism and deformation profiles under various reinforcement configurations are also studied. Geocell reinforcement is observed to take up a higher proportion of stress caused by the traffic loads compared to single and double-layer geogrid reinforcements. The efficiency of single geogrid reinforcement reduces with an increase in embedment depth. The contribution of lower geogrid is insignificant in the case of the double-geogrid reinforced system.

Keywords: Geocell, Geogrid, Flexible Pavement, Repetitive Wheel Load, Numerical Analysis

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Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

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Authors: Karl Kingsley

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Many mechanisms are involved in the control of cellular differentiation and growth, which are often dysregulated in many cancers. Many distinct pathways are involved in these mechanisms of control, including deoxyribonuclease (DNA) methyltransferase and histone deacetylase (HDAC) activation that controls both genetic and epigenetic modifications and micro ribonucleic acid (RNA) expression. Less is known about the expression of DNA methyltransferase (DNMT) and HDAC in oral cancers and the effect on microRNA expression. The primary objective of this study was to evaluate the expression of DNMT and HDAC family members in oral cancer and the concomitant expression of cancer-associated microRNAs. Using commercially available oral cancers, including squamous cell carcinoma (SCC)-4, SCC-9, SCC-15, and SCC-25, RNA was extracted and screened for DNMT, HDAC, and microRNA expression using highly-specific primers and quantitative polymerase chain reaction (qPCR). These data revealed low or absent expression of DNMT-1, which is associated with cellular differentiation but increased expression of DNMT-3a and DNMT-3b in all SCC cell lines compared with normal non-cancerous cell controls. In addition, no expression of HDAC1 and HDAC2 expression was found among the normal, non-cancerous cells but was highly expressed in each of the SCC cell lines examined. Differential expression of oncogenic and cancer-associated microRNAs was also observed among the SCC cell lines, including miR-21, miR-133, miR-149, miR-155, miR-365, and miR-720. These findings also appeared to vary according to observed growth rates among these cells. These data may be the first to demonstrate the expression and association between HDAC and DNMT3 family members among oral cancers. In addition, the differential expression of these epigenetic modifiers may be associated with the expression of specific microRNAs in these cancers, which have not previously been observed to the best of the author's knowledge. In addition, some associations and relationships may exist between the expression of these biomarkers and the rates of growth and proliferation, which may suggest that these expression patterns might represent potentially useful biomarkers to determine tumor aggressiveness and other phenotypic behaviors among oral cancers.

Keywords: oral cancer, DNA methyltransferase, histone deacetylase, microRNA

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Authors: Farusil Najeeb Mullaveettil, Rolanas Dauksevicius

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Cellular structures produced by additive manufacturing have earned wide research attention due to their unique specific strength and energy absorption potentiality. The literature review concludes that pattern type and density are vital parameters that affect the mechanical properties of parts formed by additive manufacturing techniques and have an influence on printing time and material consumption. Fused deposition modeling technique (FDM) is used here to produce Polyvinylidene fluoride (PVDF) parts. In this work, patterns are based on triply periodic minimal surfaces (TPMS) produced by PVDF-based filaments using the FDM technique. PVDF homopolymer filament Fluorinar-H™ and PVDF copolymer filament Fluorinar-C™ are printed with three types of TPMS patterns. The patterns printed are Gyroid, Schwartz diamond, and Schwartz primitive. Tensile, flexural, and compression tests under quasi-static loading conditions are performed in compliance with ISO standards. The investigation elucidates the deformation mechanisms and a study that establishes a relationship between the printed and nominal specimens' dimensional accuracy. In comparison to the examined TPMS pattern, Schwartz diamond showed a higher relative elastic modulus and strength than the other patterns in tensile loading, and the Gyroid pattern showed the highest mechanical characteristics in flexural loading. The concluded results could be utilized to produce informed cellular designs for biomedical and mechanical applications.

Keywords: additive manufacturing, FDM, PVDF, gyroid, schwartz primitive, schwartz diamond, TPMS, tensile, flexural

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Authors: Eqqab Maree, Habil Jurgen Bast, Zana K. Shakir

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Passive damping, once properly characterized and incorporated into the structure design is an autonomous mechanism. Passive damping can be achieved by applying layers of a polymeric material, called viscoelastic layers (VEM), to the base structure. This type of configuration is known as free or unconstrained layer damping treatment. A shear or constrained damping treatment uses the idea of adding a constraining layer, typically a metal, on top of the polymeric layer. Constrained treatment is a more efficient form of damping than the unconstrained damping treatment. In constrained damping treatment a sandwich is formed with the viscoelastic layer as the core. When the two outer layers experience bending, as they would if the structure was oscillating, they shear the viscoelastic layer and energy is dissipated in the form of heat. This form of energy dissipation allows the structural oscillations to attenuate much faster. The purpose behind this study is to predict damping effects by using two methods of passive viscoelastic constrained layer damping. First method is Euler-Bernoulli beam theory; it is commonly used for predicting the vibratory response of beams. Second method is Finite Element software packages provided in this research were obtained by using two-dimensional solid structural elements in ANSYS14 specifically eight nodded (SOLID183) and the output results from ANSYS 14 (SOLID183) its damped natural frequency values and mode shape for first five modes. This method of passive damping treatment is widely used for structural application in many industries like aerospace, automobile, etc. In this paper, take a steel cantilever sandwich beam with viscoelastic core type 3M-468 by using methods of passive viscoelastic constrained layer damping. Also can proved that, the percentage reduction of modal frequency between undamped and damped steel sandwich cantilever beam 8mm thickness for each mode is very high, this is due to the effect of viscoelastic layer on damped beams. Finally this types of damped sandwich steel cantilever beam with viscoelastic materials core type (3M468) is very appropriate to use in automotive industry and in many mechanical application, because has very high capability to reduce the modal vibration of structures.

Keywords: steel cantilever, sandwich beam, viscoelastic materials core type (3M468), ANSYS14, Euler-Bernoulli beam theory

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Authors: Hamid Hamli Benzahar

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The main goal of this research is to study the deflection of a composite beam CB taking into account the effect of shear deformation. The structure is made up of two beams of different sections, joined together by thin adhesive, subjected to end moments and a distributed load. The fundamental differential equation of CB can be obtained from the total energy equation while considering the shear deformation. The differential equation found will be compared with those found in CB, where the shear deformation is zero. The CB system is numerically modeled by the finite element method, where the numerical results of deflection will be compared with those found theoretically.

Keywords: composite beam, shear deformation, moments, finites elements

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Authors: A. Manallah, C. Meier

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Confocal spectral interferometry (CSI) is an innovative optical method for determining microtopography of surfaces and thickness of transparent layers, based on the combination of two optical principles: confocal imaging, and spectral interferometry. Confocal optical system images at each instant a single point of the sample. The whole surface is reconstructed by plan scanning. The interference signal generated by mixing two white-light beams is analyzed using a spectrometer. In this work, five ‘rugotests’ of known standard roughnesses are investigated. The topography is then measured and illustrated, and the equivalent roughness is determined and compared with the standard values.

Keywords: confocal spectral interferometry, nondestructive testing, optical metrology, surface topography, roughness

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Authors: Vincent Murray

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The glycopeptide bleomycin is used in the treatment of testicular cancer, Hodgkin's lymphoma, and squamous cell carcinoma. Bleomycin damages and cleaves DNA in human cells, and this is considered to be the main mode of action for bleomycin's anti-tumor activity. In particular, double-strand breaks are thought to be the main mechanism for the cellular toxicity of bleomycin. Using Illumina next-generation DNA sequencing techniques, the genome-wide sequence specificity of bleomycin-induced double-strand breaks was determined in human cells. The degree of bleomycin cleavage was also assessed at the transcription start sites (TSSs) of actively transcribed genes and compared with non-transcribed genes. It was observed that bleomycin preferentially cleaved at the TSSs of actively transcribed human genes. There was a correlation between the degree of this enhanced cleavage at TSSs and the level of transcriptional activity. Bleomycin cleavage is also affected by chromatin structure and at TSSs, the peaks of bleomycin cleavage were approximately 200 bp apart. This indicated that bleomycin was able to detect phased nucleosomes at the TSSs of actively transcribed human genes. The genome-wide cleavage pattern of the bleomycin analogues 6′-deoxy-BLM Z and zorbamycin was also investigated in human cells. As found for bleomycin, these bleomycin analogues also preferentially cleaved at the TSSs of actively transcribed human genes. The cytotoxicity (IC₅₀ values) of these bleomycin analogues was determined. It was found that the degree of enhanced cleavage at TSSs was inversely correlated with the IC₅₀ values of the bleomycin analogues. This suggested that the level of cleavage at the TSSs of actively transcribed human genes was important for the cytotoxicity of bleomycin and analogues. Hence this study provided a deeper understanding of the cellular processes involved in the cancer chemotherapeutic activity of bleomycin.

Keywords: anti-tumour activity, bleomycin analogues, chromatin structure, genome-wide study, Illumina DNA sequencing

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Authors: Thauane Silva, Gabrielle do Vale, Andre Ferreira, Marilda Siqueira, Thiago Moreno L. Souza, Milene D. Miranda

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The exposure of A(H1N1)pdm09-infected epithelial cells (HeLa) to HIV-1 viral particles, or its gp120, enhanced interferon-induced transmembrane protein (IFITM3) content, a viral restriction factor (RF), resulting in a decrease in influenza replication. The gp120 binds to CCR5 (R5) or CXCR4 (X4) cell receptors during HIV-1 infection. Then, it is possible that the endogenous ligands of these receptors also modulate the expression of IFITM3 and other cellular factors that restrict influenza virus replication. Thus, the aim of this study is to analyze the role of cellular receptors R5 and X4 in modulating RFs in order to inhibit the replication of the influenza virus. A549 cells were treated with 2x effective dose (ED50) of endogenous R5 or X4 receptor agonists, CCL3 (20 ng/ml), CCL4 (10 ng/ml), CCL5 (10 ng/ml) and CXCL12 (100 ng/mL) or exogenous agonists, gp120 Bal-R5, gp120 IIIB-X4 and its mutants (5 µg/mL). The interferon α (10 ng/mL) and oseltamivir (60 nM) were used as a control. After 24 h post agonists exposure, the cells were infected with virus influenza A(H3N2) at 2 MOI (multiplicity of infection) for 1 h. Then, 24 h post infection, the supernatant was harvested and, the viral titre was evaluated by qRT-PCR. To evaluate IFITM3 and SAM and HD domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (SAMHD1) protein levels, A549 were exposed to agonists for 24 h, and the monolayer was lysed with Laemmli buffer for western blot (WB) assay or fixed for indirect immunofluorescence (IFI) assay. In addition to this, we analyzed other RFs modulation in A549, after 24 h post agonists exposure by customized RT² Profiler Polymerase Chain Reaction Array. We also performed a functional assay in which SAMHD1-knocked-down, by single-stranded RNA (siRNA), A549 cells were infected with A(H3N2). In addition, the cells were treated with guanosine to assess the regulatory role of dNTPs by SAMHD1. We found that R5 and X4 agonists inhibited influenza replication in 54 ± 9%. We observed a four-fold increase in SAMHD1 transcripts by RFs mRNA quantification panel. After 24 h post agonists exposure, we did not observe an increase in IFITM3 protein levels through WB or IFI assays, but we observed an upregulation up to three-fold in the protein content of SAMHD1, in A549 exposed to agonists. Besides this, influenza replication enhanced in 20% in cell cultures that SAMDH1 was knockdown. Guanosine treatment in cells exposed to R5 ligands further inhibited influenza virus replication, suggesting that the inhibitory mechanism may involve the activation of the SAMHD1 deoxynucleotide triphosphohydrolase activity. Thus, our data show for the first time a direct relationship of SAMHD1 and inhibition of influenza replication, and provides perspectives for new studies on the signaling modulation, through cellular receptors, to induce proteins of great importance in the control of relevant infections for public health.

Keywords: chemokine receptors, gp120, influenza, virus restriction factors

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Authors: Jatziri Y. Moreno-Martinez, Arturo Galvan, Xavier Chavez Cardenas, Hiram Arroyo

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Several methods have been utilized to study the prediction of cracking of concrete structural under loading. The finite element analysis is an alternative that shows good results. The aim of this work was the numerical study of the width crack in reinforced concrete beams with dapped ends, these are frequently found in bridge girders and precast concrete construction. Properly restricting cracking is an important aspect of the design in dapped ends, it has been observed that the cracks that exceed the allowable widths are unacceptable in an aggressive environment for reinforcing steel. For simulating the crack width, the discrete crack approach was considered by means of a Cohesive Zone (CZM) Model using a function to represent the crack opening. Two cases of dapped-end were constructed and tested in the laboratory of Structures and Materials of Engineering Institute of UNAM. The first case considers a reinforcement based on hangers as well as on vertical and horizontal ring, the second case considers 50% of the vertical stirrups in the dapped end to the main part of the beam were replaced by an equivalent area (vertically projected) of diagonal bars under. The loading protocol consisted on applying symmetrical loading to reach the service load. The models were performed using the software package ANSYS v. 16.2. The concrete structure was modeled using three-dimensional solid elements SOLID65 capable of cracking in tension and crushing in compression. Drucker-Prager yield surface was used to include the plastic deformations. The reinforcement was introduced with smeared approach. Interface delamination was modeled by traditional fracture mechanics methods such as the nodal release technique adopting softening relationships between tractions and the separations, which in turn introduce a critical fracture energy that is also the energy required to break apart the interface surfaces. This technique is called CZM. The interface surfaces of the materials are represented by a contact elements Surface-to-Surface (CONTA173) with bonded (initial contact). The Mode I dominated bilinear CZM model assumes that the separation of the material interface is dominated by the displacement jump normal to the interface. Furthermore, the opening crack was taken into consideration according to the maximum normal contact stress, the contact gap at the completion of debonding, and the maximum equivalent tangential contact stress. The contact elements were placed in the crack re-entrant corner. To validate the proposed approach, the results obtained with the previous procedure are compared with experimental test. A good correlation between the experimental and numerical Load-Displacement curves was presented, the numerical models also allowed to obtain the load-crack width curves. In these two cases, the proposed model confirms the capability of predicting the maximum crack width, with an error of ± 30 %. Finally, the orientation of the crack is a fundamental for the prediction of crack width. The results regarding the crack width can be considered as good from the practical point view. Load-Displacement curve of the test and the location of the crack were able to obtain favorable results.

Keywords: cohesive zone model, dapped-end beams, discrete crack approach, finite element analysis

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Authors: Manuel A. Alonso-Tarajano, Roberto Mosca, Patrick Aloy

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Protein kinases participate in a myriad of cellular processes of major biomedical interest. The in vivo substrate specificity of these enzymes is a process determined by several factors, and despite several years of research on the topic, is still far from being totally understood. In the present work, we have quantified the contributions to the kinase substrate specificity of i) the phosphorylation sites and their surrounding residues in the sequence and of ii) the association of kinases to adaptor or scaffold proteins. We have used position-specific scoring matrices (PSSMs), to represent the stretches of sequences phosphorylated by 93 families of kinases. We have found negative correlations between the number of sequences from which a PSSM is generated and the statistical significance and the performance of that PSSM. Using a subset of 22 statistically significant PSSMs, we have identified specificity determinant residues (SDRs) for 86% of the corresponding kinase families. Our results suggest that different SDRs can function as positive or negative elements of substrate recognition by the different families of kinases. Additionally, we have found that human proteins with known function as adaptors or scaffolds (kAS) tend to interact with a significantly large fraction of the substrates of the kinases to which they associate. Based on this characteristic we have identified a set of 279 potential adaptors/scaffolds (pAS) for human kinases, which is enriched in Pfam domains and functional terms tightly related to the proposed function. Moreover, our results show that for 74.6% of the kinase– pAS association found, the pAS colocalize with the substrates of the kinases they are associated to. Finally, we have found evidence suggesting that the association of kinases to adaptors and scaffolds, may contribute significantly to diminish the in vivo substrate crossed- specificity of protein kinases. In general, our results indicate the relevance of several SDRs for both the positive and negative selection of phosphorylation sites by kinase families and also suggest that the association of kinases to pAS proteins may be an important factor for the localization of the enzymes with their set of substrates.

Keywords: kinase, phosphorylation, substrate specificity, adaptors, scaffolds, cellular colocalization

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Authors: Marisa Rio, Sharanya Bola, Richard H. W. Funk, Gerald Gerlach

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Cell migration, wound healing and regeneration are some of the physiological phenomena in which electric fields (EFs) have proven to have an important function. Physiologically, cells experience electrical signals in the form of transmembrane potentials, ion fluxes through protein channels as well as electric fields at their surface. As soon as a wound is created, the disruption of the epithelial layers generates an electric field of ca. 40-200 mV/mm, directing cell migration towards the wound site, starting the healing process. In vitro electrotaxis, experiments have shown cells respond to DC EFs polarizing and migrating towards one of the poles (cathode or anode). A standard electrotaxis experiment consists of an electrotaxis chamber where cells are cultured, a DC power source and agar salt bridges that help delaying toxic products from the electrodes to attain the cell surface. The electric field strengths used in such an experiment are uniform and homogeneous. In contrast, the endogenous electric field strength around a wound tend to be multi-field and non-homogeneous. In this study, we present a custom device that enables electrotaxis experiments in non-homogeneous DC electric fields. Its main feature involves the replacement of conventional metallic electrodes, separated from the electrotaxis channel by agarose gel bridges, through electrolyte-filled microchannels. The connection to the DC source is made by Ag/AgCl electrodes, incased in agarose gel and placed at the end of each microfluidic channel. An SU-8 membrane closes the fluidic channels and simultaneously serves as the single connection from each of them to the central electrotaxis chamber. The electric field distribution and current density were numerically simulated with the steady-state electric conduction module from ANSYS 16.0. Simulation data confirms the application of nonhomogeneous EF of physiological strength. To validate the biocompatibility of the device cellular viability of the photoreceptor-derived 661W cell line was accessed. The cells have not shown any signs of apoptosis, damage or detachment during stimulation. Furthermore, immunofluorescence staining, namely by vinculin and actin labelling, allowed the assessment of adhesion efficiency and orientation of the cytoskeleton, respectively. Cellular motility in the presence and absence of applied DC EFs was verified. The movement of individual cells was tracked for the duration of the experiments, confirming the EF-induced, cathodal-directed motility of the studied cell line. The in vitro monolayer wound assay, or “scratch assay” is a standard protocol to quantitatively access cell migration in vitro. It encompasses the growth of a confluent cell monolayer followed by the mechanic creation of a scratch, representing a wound. Hence, wound dynamics was monitored over time and compared for control and applied the electric field to quantify cellular population motility.

Keywords: DC non-homogeneous electric fields, electrotaxis, microfluidic biochip, wound healing

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Authors: Ozlem Oral, Seval Kilic, Ozlem Yedier, Serap Dokmeci, Devrim Gozuacik

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Autophagy is a degradation pathway, activating under stress conditions. It digests macromolecules, such as abnormal proteins and long-lived organelles by engulfing them and by subsequent delivery of the cargo to lysosomes. The members of the phospholipid-dependent serine/threonine kinases, involved in many signaling pathways, which are necessary for the regulation of cellular metabolic activation. Previous studies implicate that, serine/threonine kinases have crucial roles in the mechanism of many diseases depend on the activated and/or inactivated signaling pathway. Data indicates, the signaling pathways activated by serine/threonine kinases are also involved in activation of autophagy mechanism. However, the information about the effect of serine/threonine kinases on autophagy mechanism and the roles of these effects in disease formation is limited. In this study, we investigated the effect of activated serine/threonine kinases on autophagic pathway. We performed a commonly used autophagy technique, GFP-LC3 dot formation and by using microscopy analyses, we evaluated promotion and/or inhibition of autophagy in serine/threonine kinase-overexpressed fibroblasts as well as cancer cells. In addition, we carried out confocal microscopy analyses and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. Based on the shRNA-library based screening, we identified autophagy-related proteins affected by serine/threonine kinases. We further studied the involvement of serine/threonine kinases on the molecular mechanism of newly identified autophagy proteins and found that, autophagic pathway is indirectly controlled by serine/threonine kinases via specific autophagic proteins. Our data indicate the molecular connection between two critical cellular mechanisms, which have important roles in the formation of many disease pathologies, particularly cancer. This project is supported by TUBITAK-1001-Scientific and Technological Research Projects Funding Program, Project No: 114Z836.

Keywords: autophagy, GFP-LC3 dot formation assay, serine/threonine kinases, shRNA-library screening

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Authors: Sharmi Mukherjee, Anindita Chakraborty

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Cellular irradiation incites complex responses including arrest of cell cycle progression. This article accentuates the effects of radiation on cell cycle status of radiation non-targeted cells. Human Hepatoma HepG2 cells were exposed to increasing doses of γ radiations (1, 2, 4, 6 Gy) and their cell culture media was transferred to non-targeted HepG2 cells cultured in other Petri plates. These radiation non-targeted cells cultured in the ICCM (Irradiated cell conditioned media) were the bystander cells on which cell cycle analysis was performed using flow cytometry. An apparent decrease in the distribution of bystander cells at G0/G1 phase was observed with increased radiation doses upto 4 Gy representing a linear relationship. This was accompanied by a gradual increase in cellular distribution at G2/M phase. Interestingly the number of cells in G2/M phase at 1 and 2 Gy irradiation was not significantly different from each other. However, the percentage of G2 phase cells at 4 and 6 Gy doses were significantly higher than 2 Gy dose indicating the IC50 dose to be between 2 and 4 Gy. Cell cycle arrest is an indirect indicator of genotoxic damage in cells. In this study, bystander stress signals through the cell culture media of irradiated cells disseminated the radiation induced DNA damages in the non-targeted cells which resulted in arrest of the cell cycle progression at G2/M phase checkpoint. This implies that actual radiation biological effects represent a penumbra with effects encompassing a larger area than the actual beam. This article highlights the existence of genotoxic damages as bystander effects of γ rays in human Hepatoma cells by cell cycle analysis and opens up avenues for appraisal of bystander stress communications between tumor cells. Contemplation of underlying signaling mechanisms can be manipulated to maximize damaging effects of radiation with minimum dose and thus has therapeutic applications.

Keywords: bystander effect, cell cycle, genotoxic damage, hepatoma

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Authors: Surendra Agrawal, Pravina Gurjar, Supriya Bhide, Ram Gaud

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Levamisole hydrochloride is a prominent anticancer drug in the treatment of colon cancer but resulted in toxic effects due poor bioavailability and poor cellular uptake by tumor cells. Levamisole is an unstable drug. Incorporation of this molecule in solid lipids may minimize their exposure to the aqueous environment and partly immobilize the drug molecules within the lipid matrix-both of which may protect the encapsulated drugs against degradation. The objectives of the study were to enhance bioavailability by sustaining drug release and to reduce the toxicities associated with the therapy. Solubility of the drug was determined in different lipids to select the components of Solid Lipid Nanoparticles (SLN). Pseudoternary phase diagrams were created using aqueous titration method. Formulations were subjected to particle size and stability evaluation to select the final test formulations which were characterized for average particle size, zeta potential, and in-vitro drug release and percentage transmittance to optimize the final formulation. SLN of Levamisole hydrochloride was prepared by Nanoprecipitation method. Glyceryl behenate (Compritol 888 ATO) was used as core comprising of Tween 80 as surfactant and Lecithin as co-surfactant in (1:1) ratio. Entrapment efficiency (EE) was found to be 45.89%. Particle size was found in the range of 100-600 nm. Zeta potential of the formulation was -17.0 mV revealing the stability of the product. In-vitro release study showed that 66 % drug released in 24 hours in pH 7.2 which represent that formulation can give controlled action at the intestinal environment. In pH 5.0 it showed 64% release indicating that it can even release drug in acidic environment of tumor cells. In conclusion, results revealed SLN to be a promising approach to sustain the drug release so as to increase bioavailability and cellular uptake of the drug with reduction in toxic effects as dose has been reduced with controlled delivery.

Keywords: SLN, nanoparticulate delivery of levamisole, pharmacy, pharmaceutical sciences

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Authors: Franco Primo Soffietti, Diego Fernando Turello, Federico Pinto

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Shear waves that propagate through the ground impose deformations that must be taken into account in the design and assessment of buried longitudinal structures such as tunnels, pipelines, and piles. Conventional engineering approaches for seismic evaluation often rely on a Euler-Bernoulli beam models supported by a Winkler foundation. This approach, however, falls short in capturing the distortions induced when the structure is subjected to shear waves. To overcome these limitations, in the present work an analytical solution is proposed considering a Timoshenko beam and including transverse and rotational springs. The present research proposes ground springs derived as closed-form analytical solutions of the equations of elasticity including the seismic wavelength. These proposed springs extend the applicability of previous plane-strain models. By considering variations in displacements along the longitudinal direction, the presented approach ensures the springs do not approach zero at low frequencies. This characteristic makes them suitable for assessing pseudo-static cases, which typically govern structural forces in kinematic interaction analyses. The results obtained, validated against existing literature and a 3D Finite Element model, reveal several key insights: i) the cutoff frequency significantly influences transverse and rotational springs; ii) neglecting displacement variations along the structure axis (i.e., assuming plane-strain deformation) results in unrealistically low transverse springs, particularly for wavelengths shorter than the structure length; iii) disregarding lateral displacement components in rotational springs and neglecting variations along the structure axis leads to inaccurately low spring values, misrepresenting interaction phenomena; iv) transverse springs exhibit a notable drop in resonance frequency, followed by increasing damping as frequency rises; v) rotational springs show minor frequency-dependent variations, with radiation damping occurring beyond resonance frequencies, starting from negative values. This comprehensive analysis sheds light on the complex behavior of embedded longitudinal structures when subjected to shear waves and provides valuable insights for the seismic assessment.

Keywords: shear waves, Timoshenko beams, Winkler springs, sol-structure interaction

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Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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In this study, in the breeding Japanese quails (coturnix coturnix japonica), it was aimed to study the effects of cellular insulin receptor stimulation on the performance, some blood parameters, and hatchability features. In the study, a total of 84 breeding quails were used, which are in 6 weeks age, and whose 24 are male and 60 female. In the trial, rations which contain 2900 kcal/kg metabolic energy; crude protein of 20%, and water whose pH is calibrated to 7.45 were administered as ad-libitum, to the animals, as metformin source, metformin-HCl was used and as chrome resource, Chromium Picolinate. Trial groups were formed as control group (basal ration), metformin group (basal ration, added metformin at the level of fodder of 20 mg/kg), and chromium picolinate group (basal ration, added fodder of 1500 ppb Cr. When regarded to the results of performance at the end of trial, it is seen that live weight gain, fodder consumption, egg weight, fodder evaluation coefficient, and egg production were affected at the significant level (p < 0.05). When the results are evaluated in terms of incubation features at the end of trial, it was identified that incubation yield and hatchability are not affected by the treatments but in the groups, in which metformin and chromium picolinate are added to ration, that fertility rose at the significant level compared to control group (p < 0,05). According to the results of blood parameters and hormone at the end of the trial, while the level of plasma glucose level was not affected by treatments (p > 0.05), with the addition of metformin and chromium picolinate to ration, plasma, total control, cholesterol, HDL, LDL, and triglyceride levels were significantly affected from insulin receptor stimulators added to ration (p<0,05). Hormone level of Plasma T3 and T4 were also affected at the significant level from insulin receptor stimulators added to ration (p < 0,05).

Keywords: cholesterol, chromium picolinate, hormone, metformin, performance, quail

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Authors: Meisam Omidi, M. Mirijalili, Mohammadmehdi Choolaei, Z. Sharifi, F. Haghiralsadat, F. Yazdian

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In this work, we have used arrays of micromechanical piezoresistive cantilever with different geometries to detect carcinoembryonic antigen (CEA), which is known as an important biomarker associated with various cancers such as the colorectal, lung, breast, pancreatic, and bladder cancer. The sensing principle is based on the surface stress changes induced by antigen–antibody interaction on the microcantilevers surfaces. Different concentrations of CEA in a human serum albumin (HSA) solution were detected as a function of the deflection of the beams. According to the experiments, it was revealed that microcantilevers have surface stress sensitivities in the order of 8 (mJ/m). This matter allows them to detect CEA concentrations as low as 3 ng/mL or 18 pM. This indicates the fact that the self-sensing microcantilever approach is beneficial for pathological tests.

Keywords: micromechanical biosensors, carcinoembryonic antigen (CEA), surface stress

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Authors: M. Li, V. D. Thi, M. Khelifa, M. El Ganaoui

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This study presents an overview of the work carried out by the use of wood waste as coarse aggregate in mortar. The paper describes experimental and numerical investigations carried on pervious concrete made of wood chips and also sheds lights on the mechanical properties of this new product. The properties of pervious wood-concrete such as strength, elastic modulus, and failure modes are compared and evaluated. The characterization procedure of the mechanical properties of wood waste ash are presented and discussed. The numerical and tested load–deflection response results are compared. It was observed that the numerical results are in good agreement with the experimental results.

Keywords: wood waste ash, characterization, mechanical properties, bending tests

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Authors: Marcelo Sanhueza Cartes, Nelson Maureira Carsalade

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This research evaluates the effectiveness of a pallet isolation device for the protection of selective-type industrial storage racks. The device works only in the longitudinal direction of the aisle, and it is made up of a platform installed on the rack beams. At both ends, the platform is connected to the rack structure by means of a spring-damper system working in parallel. A system of wheels is arranged between the isolation platform and the rack beams in order to reduce friction, decoupling of the movement and improve the effectiveness of the device. The latter is evaluated by the reduction of the maximum dynamic responses of basal shear load and story drift in relation to those corresponding to the same rack with the traditional construction system. In the first stage, numerical simulations of industrial storage racks were carried out with and without the pallet isolation device. The numerical results allowed us to identify the archetypes in which it would be more appropriate to carry out experimental tests, thus limiting the number of trials. In the second stage, experimental tests were carried out on a shaking table to a select group of full-scale racks with and without the proposed device. The movement simulated by the shaking table was based on the Mw 8.8 magnitude earthquake of February 27, 2010, in Chile, registered at the San Pedro de la Paz station. The peak ground acceleration (PGA) was scaled in the frequency domain to fit its response spectrum with the design spectrum of NCh433. The experimental setup contemplates the installation of sensors to measure relative displacement and absolute acceleration. The movement of the shaking table with respect to the ground, the inter-story drift of the rack and the pallets with respect to the rack structure were recorded. Accelerometers redundantly measured all of the above in order to corroborate measurements and adequately capture low and high-frequency vibrations, whereas displacement and acceleration sensors are respectively more reliable. The numerical and experimental results allowed us to identify that the pallet isolation period is the variable with the greatest influence on the dynamic responses considered. It was also possible to identify that the proposed device significantly reduces both the basal cut and the maximum inter-story drift by up to one order of magnitude.

Keywords: pallet isolation system, industrial storage racks, basal shear load, interstory drift.

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Authors: Yaw Opoku-Damoah, Run Zhang, Hang Thu Ta, Zhi Ping Xu

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Gas therapy is still at an early stage of research and development. Even though most gasotransmitters have proven their therapeutic potential, their handling, delivery, and controlled release have been extremely challenging. This research work employs a versatile nanosystem that is capable of delivering a gasotransmitter in the form of a photo-responsive carbon monoxide-releasing molecule (CORM) for targeted cancer therapy. The therapeutic action was mediated by upconversion nanoparticles (UCNPs) designed to transfer bio-friendly low energy near-infrared (NIR) light to ultraviolet (UV) light capable of triggering carbon monoxide (CO) from a water-soluble amphiphilic manganese carbonyl complex CORM incorporated into a carefully designed lipid drug delivery system. Herein, gaseous CO that plays a role as a gasotransmitter with cytotoxic and homeostatic properties was investigated to instigate cellular apoptosis. After successfully synthesizing the drug delivery system, the ability of the system to encapsulate and mediate the sustained release of CO after light excitation was demonstrated. CO fluorescence probe (COFP) was successfully employed to determine the in vitro drug release profile upon NIR light irradiation. The uptake of nanoparticles enhanced by folates and its receptor interaction was also studied for cellular uptake purposes. The anticancer potential of the final lipid nanoparticle Lipid/UCNPs/CORM/FA (LUCF) was also determined by cell viability assay. Intracellular CO release and a subsequent therapeutic action involving ROS production, mitochondrial damage, and CO production was also evaluated. In all, this current project aims to use in vitro studies to determine the potency and efficiency of a NIR-mediated CORM prodrug delivery system.

Keywords: carbon monoxide-releasing molecule, upconversion nanoparticles, site-specific delivery, amphiphilic manganese carbonyl complex, prodrug delivery system.

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Authors: Anil Koklu, Amin Mansoorifar, Ali Beskok

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Dielectric spectroscopy (DS) is a noninvasive, label free technique for a long term real-time measurements of the impedance spectra of biological cells. DS enables characterization of cellular dielectric properties such as membrane capacitance and cytoplasmic conductivity. We have developed a lab-on-a-chip device that uses an electro-activated microwells array for loading, DS measurements, and unloading of biological cells. We utilized from dielectrophoresis (DEP) to capture target cells inside the wells and release them after DS measurement. DEP is a label-free technique that exploits differences among dielectric properties of the particles. In detail, DEP is the motion of polarizable particles suspended in an ionic solution and subjected to a spatially non-uniform external electric field. To the best of our knowledge, this is the first microfluidic chip that combines DEP and DS to analyze biological cells using electro-activated wells. Device performance is tested using two different cell lines of prostate cancer cells (RV122, PC-3). Impedance measurements were conducted at 0.2 V in the 10 kHz to 40 MHz range with 6 s time resolution. An equivalent circuit model was developed to extract the cell membrane capacitance and cell cytoplasmic conductivity from the impedance spectra. We report the time course of the variations in dielectric properties of PC-3 and RV122 cells suspended in low conductivity medium (LCB), which enhances dielectrophoretic and impedance responses, and their response to sudden pH change from a pH of 7.3 to a pH of 5.8. It is shown that microfluidic chip allowed online measurements of dielectric properties of prostate cancer cells and the assessment of the cellular level variations under external stimuli such as different buffer conductivity and pH. Based on these data, we intend to deploy the current device for single cell measurements by fabricating separately addressable N × N electrode platforms. Such a device will allow time-dependent dielectric response measurements for individual cells with the ability of selectively releasing them using negative-DEP and pressure driven flow.

Keywords: microfluidic, microfabrication, lab on a chip, AC electrokinetics, dielectric spectroscopy

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Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

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Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

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Authors: Zeina Merabi, Arij Dao

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The temporal order of information processing in the brain is an important code in many acoustic signals, including speech, music, and animal vocalizations. Despite its significance, surprisingly little is known about its underlying cellular mechanisms and network manifestations. In the songbird telencephalic nucleus HVC, a subset of neurons shows temporal combination sensitivity (TCS). These neurons show a high temporal specificity, responding differently to distinct patterns of spectral elements and their combinations. HVC neuron types include basal-ganglia-projecting HVCX, forebrain-projecting HVCRA, and interneurons (HVC¬INT), each exhibiting distinct cellular, electrophysiological and functional properties. In this work, we develop conductance-based neural network models connecting the different classes of HVC neurons via different wiring scenarios, aiming to explore possible neural mechanisms that orchestrate the combination sensitivity property exhibited by HVCX, as well as replicating in vivo firing patterns observed when TCS neurons are presented with various auditory stimuli. The ionic and synaptic currents for each class of neurons that are presented in our networks and are based on pharmacological studies, rendering our networks biologically plausible. We present for the first time several realistic scenarios in which the different types of HVC neurons can interact to produce this behavior. The different networks highlight neural mechanisms that could potentially help to explain some aspects of combination sensitivity, including 1) interplay between inhibitory interneurons’ activity and the post inhibitory firing of the HVCX neurons enabled by T-type Ca2+ and H currents, 2) temporal summation of synaptic inputs at the TCS site of opposing signals that are time-and frequency- dependent, and 3) reciprocal inhibitory and excitatory loops as a potent mechanism to encode information over many milliseconds. The result is a plausible network model characterizing auditory processing in HVC. Our next step is to test the predictions of the model.

Keywords: combination sensitivity, songbirds, neural networks, spatiotemporal integration

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Authors: Mehrnaz Mostafavi

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Lung cancer is one of the most harmful forms of cancer. The long-term survival rate of lung cancer patients treated by conventional modalities such as surgical resection, radiation, and chemotherapy remains far from satisfactory. Systemic drug delivery is rarely successful because only a limited amount of the chemotherapeutic drug targets lung tumor sites, even when administered at a high dose. Targeted delivery of drug molecules to organs or special sites is one of the most challenging research areas in pharmaceutical sciences. By developing colloidal delivery systems such as liposomes, micelles and nanoparticles a new frontier was opened for improving drug delivery. Nanoparticles with their special characteristics such as small particle size, large surface area and the capability of changing their surface properties have numerous advantages compared with other delivery systems. Targeted nanoparticle delivery to the lungs is an emerging area of interest.Multimodal or combination therapy represents a promising new method to fight disease. Therefore, a combination of different therapeutic strategies may be the best alternative to improve treatment outcomes for lung cancer. Photothermal therapy was proposed as a novel approach to treatment. In this work, photothermal therapy with gold nanoparticles and near infrared laser (NIR) irradiation was investigated.Four types of small (<100nm), NIR absorbing gold nanoparticles (nanospheres, nanorods) were synthesized using wet chemical methods and characterized by transmission electron microscopy, dynamic light scattering and UV-vis spectroscopy. Their synthesis and properties were evaluated, to determine their feasibility as a photothermal agent for clinical applications. In vitro cellular uptake studies of the nanoparticles into lung cancer cell lines was measured using light scattering microscopy.Small gold nanorods had good photothermal properties and the greatest cellular uptake, and were used in photothermal studies. Under 4W laser irradiation, an increase in temperature of 10°C and decrease in cell viability of up to 80% were obtained.

Keywords: photothermal, therapy, cancer, gold nanorods

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Authors: Shabnam Abdi, Behzad Toosi

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Background: Melanoma is the most lethal type of malignant skin cancer in humans and dogs since it spreads rapidly throughout the body. Despite significant advances in treatment, cancer at an advanced stage has a poor prognosis. Hence, more effective treatments are needed to enhance outcomes with fewer side effects. Erythropoietin-producing hepatocellular receptors are the largest family of receptor tyrosine kinases and are divided into two subfamilies, EphA and EphB, both of which play a significant role in disease, especially cancer. Due to their association with proliferation and invasion in many aggressive types of cancer, Eph receptor tyrosine kinases (Eph RTKs) are promising cancer therapy molecules. Because these receptors have not been studied in canine melanoma, we investigated how EphA2 influences survival and tumorigenicity of melanoma cells. Methods: Expression of EphA2 protein in canine melanoma cell lines and human melanoma cell line was evaluated by Western blot. Melanoma cells were transduced with lentiviral particles encoding Eph-targeting shRNAs or non-silencing shRNAs (control) for silencing the expression of EphA2 receptor, and silencing was confirmed by Western blotting and immunofluorescence. The effect of siRNA treatment on cellular proliferation, colony formation, tumorsphere assay, invasion was analyzed by Resazurin assay Matrigel invasion assay, respectively. Results: Expression of EphA2 was detected in canine and human melanoma cell lines. Moreover, stably silencing EphA2 by specific shRNAs significantly and consistently decreased the expression of EphA2 protein in both human and canine melanoma cells. Proliferation, colony formation, tumorsphere and invasion of melanoma cells were significantly decreased in EphA2 siRNA-treated cells compared to control. Conclusion: Our data provide the first functional evidence that the EphA2 receptor plays a critical role in the malignant cellular behavior of melanoma in both human and dogs.

Keywords: ephA2, targeting, melanoma, human, canine

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Authors: Morteza Raki, Abolghasem Zabihollah, Omid Askari

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Cantilever beam is a simplified sample of a lot of mechanical components used in a wide range of applications, including many industries such as gas turbine blade. Due to the nature of the operating conditions, beams are subject to variety of damages especially crack propagates. Crack propagation may lead to catastrophic failure during operation. Therefore, online detection of crack presence and its propagation is very important and may reduce possible significant cost of the whole system failure. This paper aims to investigate the effect of cracks presence and crack propagation on one end fixed beam`s vibration. A finite element model will be developed for the blade in which the modal response of the structure with and without crack will be studied. 

Keywords: blade, crack propagation, health monitoring, modal analysis

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

Abstract:

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

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

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Authors: Dimakatso B. Gumede, Nicolette N. Houreld

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Diabetic foot ulcers (DFUs) are a complication of diabetes mellitus (DM), a metabolic disease caused by insulin resistance or insufficiency, resulting in hyperglycaemia and low-grade chronic inflammation. Current therapies for treating DFUs include wound debridement, glycaemic control, and wound dressing. However, these therapies are moderately effective as there is a recurrence of these ulcers and an increased risk of lower limb amputations. Photobiomodulation (PBM), which is the application of non-invasive low-level light for wound healing at the spectrum of 660-1000 nm, has shown great promise in accelerating the healing of chronic wounds. However, its underlying mechanisms are not clearly defined. Studies have indicated that PBM induces wound healing via the activation of signaling pathways that are involved in tissue repair, such as the transforming growth factor-β (TGF-β). However, other signaling pathways, such as the WNT/β-catenin pathway, which is also critical for wound repair, have not been investigated. This study aimed to elucidate if PBM at 660 nm and a fluence of 5 J/cm² activates the WNT/β-catenin signaling pathway for wound healing in a diabetic cellular model. Human dermal fibroblasts (WS1) were continuously cultured high-glucose (26.5 mM D-glucose) environment to create a diabetic cellular model. A central scratch was created in the diabetic model to ‘wound’ the cells. The diabetic wounded (DW) cells were thereafter irradiated at 660 nm and a fluence of 5 J/cm². Cell migration, gene expression and protein assays were conducted at 24- and 48-h post-PBM. The results showed that PBM at 660 nm and a fluence of 5 J/cm² significantly increased cell migration in diabetic wounded cells at 24-h post-PBM. The expression of CTNNB1, ACTA2, COL1A1 and COL3A1 genes was also increased in DW cells post-PBM. Furthermore, there was increased cytoplasmic accumulation and nuclear localization of β-catenin at 24 h post-PBM. The findings in this study demonstrate that PBM activates the WNT/β-catenin signaling pathway by inducing the accumulation of β-catenin in diabetic wounded cells, leading to increased cell migration and expression of wound repair markers. These results thus indicate that PBM has the potential to improve wound healing in diabetic ulcers via activation of the WNT/β-catenin signaling pathway.

Keywords: wound healing, diabetic ulcers, photobiomodulation, WNT/β-catenin, signalling pathway

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