Search results for: in vitro activation

Authors: Biswabandhu Bankura, Srikanta Guria, Madhusudan Das

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Purpose: Thyroid peroxidase (TPO) is the key enzyme in the biosynthesis of thyroid hormones. We aimed to identify the spectrum of mutations in the TPO gene leading to hypothyroidism in the population of West Bengal to establish the genetic etiology of the disease. Methods: 200 hypothyroid patients (case) and their corresponding sex and age matched 200 normal individuals (control) were screened depending on their clinical manifestations. Genomic DNA was isolated from peripheral blood samples and TPO gene (Exon 7 to Exon 14) was amplified by PCR. The PCR products were subjected to sequencing to identify mutations. Results: Single nucleotide changes such as Glu 641 Lys, Asp 668 Asn, Thr 725 Pro, Asp 620 Asn, Ser 398 Thr, and Ala 373 Ser were found. Changes in the TPO were assayed in vitro to compare mutant and wild-type activities. Five mutants were enzymatically inactive in the guaiacol and iodide assays. This is a strong indication that the mutations are present at crucial positions of the TPO gene, resulting in inactivated TPO. Key Findings: The results of this study may help to develop a genetic screening protocol for goiter and hypothyroidism in the population of West Bengal.

Keywords: thyroid peroxidase, hypothyroidism, mutation, in vitro assay, transfection

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Authors: Jiahui Song, Ravindra Joshi

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High-intensity, nanosecond, pulsed electric fields have been shown to be useful non-thermal tools capable of producing a variety of specific cellular responses. While reversible and temporary changes are often desired based on electromanipulation, irreversible effects can also be important objectives. These include elimination of tumor cells and bacterial decontamination. A simple model-based rate-equation treatment of the various cellular biochemical processes was used to qualitatively predict the pulse number-dependent caspase activation and cell survival trends. The model incorporated the caspase-8 associated extrinsic pathway, the delay inherent in its activation, cytochrome c release, and the internal feedback mechanism between caspase-3 and Bid. Results were roughly in keeping with the experimental cell-survival data. A pulse-number threshold was predicted followed by a near-exponential fall-off. The intrinsic pathway was shown to be much weaker as compared to the extrinsic mechanism for electric pulse induced cell apoptosis. Also, delays of about an hour are predicted for detectable molecular concentration increases following electrical pulsing.

Keywords: apoptosis, cell survival, model, pathway

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Authors: Steven Arcidiacono, Robert Stote, Erin Anderson, Molly Richards

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There are numerous standard test methods for antimicrobial textiles that measure activity against specific microorganisms. However, many times these results do not translate to the performance of treated textiles when worn by individuals. Standard test methods apply a single target organism grown under optimal conditions to a textile, then recover the organism to quantitate and determine activity; this does not reflect the actual performance environment that consists of polymicrobial communities in less than optimal conditions or interaction of the textile with the skin substrate. Here we propose the development of in vitro skin model method to bridge the gap between lab testing and wear studies. The model will consist of a defined polymicrobial community of 5-7 commensal microbes simulating the skin microbiome, seeded onto a solid tissue platform to represent the skin. The protocol would entail adding a non-commensal test organism of interest to the defined community and applying a textile sample to the solid substrate. Following incubation, the textile would be removed and the organisms recovered, which would then be quantitated to determine antimicrobial activity. Important parameters to consider include identification and assembly of the defined polymicrobial community, growth conditions to allow the establishment of a stable community, and choice of skin surrogate. This model could answer the following questions: 1) is the treated textile effective against the target organism? 2) How is the defined community affected? And 3) does the textile cause unwanted effects toward the skin simulant? The proposed model would determine activity under conditions comparable to the intended application and provide expanded knowledge relative to current test methods.

Keywords: antimicrobial textiles, defined polymicrobial community, in vitro skin model, skin microbiome

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Authors: Yuvraj Singh Dangi, Kamta Prasad Namdeo, Surendra Bodhake

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The purpose of this research was to develop and evaluate mucoadhesive films for buccal administration of itraconazole using film-forming and mucoashesive polymers. Buccal films of chitosan bearing Itraconazole were prepared by solvent casting technique. The films have been evaluated in terms of film weight, thickness, density, surface pH, FTIR, X-ray diffraction analysis, bioadhesion, swelling properties, and in vitro drug release studies. It was found that film formulations of 2 cm2 size having weight in the range of 204 ± 0.76 to 223 ± 2.09 mg and film thickness were in the range of 0.44 ± 0.11 to 0.57 ± 0.19 mm. Density of the films was found to be 0.102 to 0.126 g/ml. Drug content was found to be uniform in the range of 8.23 ± 0.07 to 8.73 ± 0.09 mg/cm2 for formulation A1 to A4. Maximum bioadhesion force was recorded for HPMC buccal films (A2) i.e. 0.57 ± 0.47 as compared to other films. In vitro residence time was in range of 1.7 ± 0.12 to 7.65 ± 0.15 h. The drug release studies show that formulations follow non-fickian diffusion. These mucoadhesive formulations could offer many advantages in comparison to traditional treatments.

Keywords: biovariability, buccal patches, itraconazole, Mucoadhesion

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

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

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

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Authors: Marta Błażkiewicz, Adam Konefał

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Currently, boron-neutron therapy is carried out mainly with the use of a neutron beam generated in research nuclear reactors. This fact limits the possibility of realization of a BNCT in centers distant from the above-mentioned reactors. Moreover, the number of active nuclear reactors in operation in the world is decreasing due to the limited lifetime of their operation and the lack of new installations. Therefore, the possibilities of carrying out boron-neutron therapy based on the neutron beam from the experimental reactor are shrinking. However, the use of nuclear power reactors for BNCT purposes is impossible due to the infrastructure not intended for radiotherapy. Therefore, a serious challenge is to find ways to perform boron-neutron therapy based on neutrons generated outside the research nuclear reactor. This work meets this challenge. Its goal is to develop a BNCT technique based on commonly available neutron sources such as Cf-252 and PuBe, which will enable the above-mentioned therapy in medical centers unrelated to nuclear research reactors. Advances in the field of neutron source fabrication make it possible to achieve strong neutron fluxes. The current stage of research focuses on the development of virtual models of the above-mentioned sources using the Monte Carlo simulation method. In this study, the GEANT4 tool was used, including the model for simulating neutron-matter interactions - High Precision Neutron. Models of neutron sources were developed on the basis of experimental verification based on the activation detectors method with the use of indium foil and the cadmium differentiation method allowing to separate the indium activation contribution from thermal and resonance neutrons. Due to the large number of factors affecting the result of the verification experiment, the 10% discrepancy between the simulation and experiment results was accepted.

Keywords: BNCT, virtual models, neutron sources, monte carlo, GEANT4, neutron activation detectors, gamma spectroscopy

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Authors: Wenxing Chen, Guangying Chen, Yin Lu, Shile Huang

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Cryptotanshinone (CPT), a fat-soluble tanshinone from Salvia miltiorrhiza Bunge, has been demonstrated to inhibit mTOR pathway, resulting in inhibition of cancer cell proliferation. However, the molecular mechanism how CPT acts on mTOR is unknown. Here, cancer cells expressing rapamycin-resistant mutant mTOR are also sensitive to CPT, while phosphorylation of AMPK and TSC2 was activated, suggesting that CPT inhibition of mTOR maybe due to activating upstream of mTOR, AMPK, but not directly binding to and inhibiting mTOR. Further results indicated that Compound C, inhibitor of AMPK, could partially reversed CPT inhibition effect on cancer cells, and dominant-negative AMPK in cancer cells conferred resistance to CPT inhibition of 4EBP1 and phosphorylation of S6K1, as well as sh-AMPK. Furthermore, compared with MEF cells with AMPK positive, MEF cells with AMPK knock out are less sensitive to CPT by the findings that 4E-BP1 and phosphorylation of S6K1 express comparatively much. Furthermore, downexpression of TSC2 slightly recovered expression of 4EBP1 and phosphorylation of S6K1, while co-immunoprecipitation of TSC2 did not affect expression of TSC1 by CPT. Collectively, the above-mentioned results suggest that CPT inhibited mTOR pathway mostly was due to activation of AMPK-TSC2 pathway rather than specific inhibition of mTOR and then induction of subsequent lethal cellular effect.

Keywords: cryptotanshinone, AMPK, TSC2, mTOR, cancer cells

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Authors: Woolfrey K., Woolfrey M., Bolton C. L., Warchuk D.

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Objectives: Osteoarthritis is the most common joint disease of adults worldwide. Despite total knee arthroplasty (TKA) demonstrating high levels of success, 20% of patients report dissatisfaction with their result. VOXX Wellness Stasis Socks are embedded with a proprietary pattern of neuro-receptor activation points that have been proven to activate a precise neuro-response, according to the pattern theory of haptic perception, which stimulates improvements in pain and function. The use of this technology in TKA patients may prove beneficial as an adjunct to recovery as many patients suffer from deficits to their proprioceptive system caused by ligamentous damage and alterations to mechanoreceptors during the procedure. We hypothesized that VOXX Wellness Stasis Socks are a safe, cost-effective, and easily scalable strategy to support TKA patients through their recovery. Design: Double-blinded, placebo-controlled randomized trial. Participants: Patients scheduled to receive TKA were considered eligible for inclusion in the trial. Interventions: Intervention group (I): VOXX Wellness Stasis socks containing receptor point-activation technology. Control group (C): VOXX Wellness Stasis socks without receptor point-activation technology. Sock use during the waking hours x 6 weeks. Main Outcome Measures: Western Ontario McMaster Universities Osteoarthritis Index (WOMAC) questionnaire completed at baseline, 2 weeks, and 6 weeks to assess pain, stiffness, and physical function. Results: Data analysis using SPSS software. P-values, effect sizes, and confidence intervals are reported to assess clinical relevance of the finding. Physical status classifications were compared using t-test. Within-subject and between-subject differences in the mean WOMAC were analyzed by ANOVA. Effect size was analyzed using Cramer’s V. Consistent improvement in WOMAC scores for pain and stiffness at 2 weeks post op in the I over the C group. The womac scores assessing physical function showed a consistent improvement at both 2 and 6 weeks post op in the I group compared to C group. Conclusions: VOXX proved to be a low cost, safe intervention in TKA to help patients improve with regard to pain, stiffness, and physical function. Disclosures: None

Keywords: osteoarthritis, RCT, pain management, total knee arthroplasty

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Authors: Farhan Sohail, Gul Shahnaz Irshad Hussain, Shoaib Sarwar, Ibrahim Javed, Zajif Hussain, Akhtar Nadhman

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The present study was aimed to develop a hybrid nanocarrier (NC) system with enhanced membrane permeability, bioavailability and targeted delivery of Docetaxel (DTX) in breast cancer. Hybrid NC’s based on folic acid (FA) grafted thiolated chitosan (TCS) enveloped liposomes were prepared with DTX and evaluated in-vitro and in-vivo for their enhanced permeability and bioavailability. Physicochemical characterization of NC’s including particle size, morphology, zeta potential, FTIR, DSC, PXRD, encapsulation efficiency and drug release from NC’s was determined in vitro. Permeation enhancement and p-gp inhibition were performed through everted sac method on freshly excised rat intestine which indicated that permeation was enhanced 5 times as compared to pure DTX and the hybrid NC’s were strongly able to inhibit the p-gp activity as well. In-vitro cytotoxicity and tumor targeting was done using MDA-MB-231 cell line. The stability study of the formulations performed for 3 months showed the improved stability of FA-TCS enveloped liposomes in terms of its particles size, zeta potential and encapsulation efficiency as compared to TCS NP’s and liposomes. The pharmacokinetic study was performed in vivo using rabbits. The oral bioavailability and AUC0-96 was increased 10.07 folds with hybrid NC’s as compared to positive control. Half-life (t1/2) was increased 4 times (58.76 hrs) as compared to positive control (17.72 hrs). Conclusively, it is suggested that FA-TCS enveloped liposomes have strong potential to enhance permeability and bioavailability of hydrophobic drugs after oral administration and tumor targeting.

Keywords: docetaxel, coated liposome, permeation enhancement, oral bioavailability

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Authors: Kiran Nawaz, Ahmad Ali Shahid, Sehrish Iftikhar, Waheed Anwar, Muhammad Nasir Subhani

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Chili (Solanum annum L.) attacks by many fungal pathogens, including members of Oomycetes which are responsible for root rot in different chili growing areas of the world. Oomycetes pathogens cause economic losses in different regions of the Pakistan. Most of the plant tissues, including roots, crowns, fruit, and leaves, are vulnerable to Phytophthora capsici. It is very difficult to manage the Phytophthora root rot of chili as many commercial varieties are tremendously vulnerable to P. capsici. The causal agent of the disease was isolated on corn meal agar (CMA) and identified on a morphological basis by using available taxonomic keys. The pathogen was also confirmed on the molecular basis through internal transcribed spacer region and with other molecular markers.The Blastn results showed 100% homology with already reported sequences of P. capsici in NCBI database. Most of the farmers have conventionally relied on foliar fungicide applications to control Phytophthora root rot in spite of their incomplete effectiveness. In this study, in vitro plate assay, seed soaking and foliar applications of 6 fungicides were evaluated against root rot of chili. In vitro assay revealed that significant inhibition of linear growth was obtained with Triflumizole at 7.0%, followed by Thiophanate methyl (8.9%), Etridiazole (6.0%), Propamocarb (5.9%) and 7.5% with Mefenoxam and Iprodione for P. capsici. The promising treatments of in vitro plate bioassay were evaluated in pot experiments under controlled conditions in the greenhouse. All fungicides were applied after at 6-day intervals. Results of pot experiment showed that all treatments considerably inhibited the percentage of P. capsici root rot incidence. In addition, application of seed soaking with all six fungicides combined with the foliar spray of the same components showed the significant reduction in root rot incidence. The combine treatments of all fungicides as in vitro bioassay, seed soaking followed by foliar spray is considered non-harmful control methods which have advantages and limitation. Hence, these applications proved effective and harmless for the management of soil-borne plant pathogens.

Keywords: blastn, bioassay, corn meal agar(CMA), oomycetes

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Authors: Marcela Capcarova, Adriana Kolesarova, Marina Medvedova, Peter Petruska, Alexander V. Sirotkin

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The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.

Keywords: deoxynivalenol, zearalenone, antioxidants, Hsp70, granulosa cells

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Authors: Jessica Arthur, Duke Amegah, Kingsley Akenten Wiafe

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Background: Vanilla planifolia is the only edible fruit of the orchid family (Orchidaceae) among the over 35,000 Orchidaceae species found worldwide. In Ghana, Vanilla was discovered in the wild, but it is underutilized for commercial production, most likely due to a lack of knowledge on the best NAA and BAP combinations for in vitro propagation to promote successfully regenerated plant acclimatization. The growing interest and global demand for elite Vanilla planifolia plants and natural vanilla flavour emphasize the need for an effective industrial-scale micropropagation protocol. Tissue culture systems are increasingly used to grow disease-free plants and reliable in vitro methods can also produce plantlets with typically modest proliferation rates. This study sought to develop an efficient protocol for in vitro propagation of vanilla using nodal explants by testing different concentrations of NAA and BAP, for the proliferation of the entire plant. Methods: Nodal explants with dormant axillary buds were obtained from year-old laboratory-grown Vanilla planifolia plants. MS media was prepared with a nutrient stock solution (containing macronutrients, micronutrients, iron solution and vitamins) and semi-solidified using phytagel. It was supplemented with different concentrations of NAA and BAP to induce multiple shoots and roots (0.5mg/L BAP with NAA at 0, 0.5, 1, 1.5, 2.0mg/L and vice-versa). The explants were sterilized, cultured in labelled test tubes and incubated at 26°C ± 2°C with 16/8 hours light/dark cycle. Data on shoot and root growth, leaf number, node number, and survival percentage were collected over three consecutive two-week periods. The data were square root transformed and subjected to ANOVA and LSD at a 5% significance level using the R statistical package. Results: Shoots emerged at 8 days and roots at 12 days after inoculation with 94% survival rate. It was discovered that for the NAA treatments, MS media supplemented with 2.00 mg/l NAA resulted in the highest shoot length (10.45cm), maximum root number (1.51), maximum shoot number (1.47) and the highest number of leaves (1.29). MS medium containing 1.00 mg/l NAA produced the highest number of nodes (1.62) and root length (14.27cm). Also, a similar growth pattern for the BAP treatments was observed. MS medium supplemented with 1.50 mg/l BAP resulted in the highest shoot length (14.98 cm), the highest number of nodes (4.60), the highest number of leaves (1.75) and the maximum shoot number (1.57). MS medium containing 0.50 mg/l BAP and 1.0 mg/l BAP generated a maximum root number (1.44) and the highest root length (13.25cm), respectively. However, the best concentration combination for maximizing shoot and root was media containing 1.5mg/l BAP combined with 0.5mg/l NAA, and 1.0mg/l NAA combined with 0.5mg/l of BAP respectively. These concentrations were optimum for in vitro growth and production of Vanilla planifolia. Significance: This study presents a standardized protocol for labs to produce clean vanilla plantlets, enhancing cultivation in Ghana and beyond. It provides insights into Vanilla planifolia's growth patterns and hormone responses, aiding future research and cultivation.

Keywords: Vanilla planifolia, In vitro propagation, plant hormones, MS media

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Authors: V. Miller, T. Jariel, C. Cooper-Chadwick

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Early treatment of stroke leads to higher survival and lower disability rates. Increasing knowledge to activate the emergency medical system for signs of stroke can improve outcomes for patients with stroke and decrease morbidity and mortality. Even though patients who get discharged from the hospital receive standard verbal and printed education, nearly 20% of them answer the question incorrectly when asked, “What will you do if you or someone you know have signs of stroke?” The main goal of this evidence-based project was to improve patients’ knowledge of what to do if they have signs of stroke. Evidence suggests that using video education in conjunction with verbal and printed education improves patient comprehension and retention. The percentage of patients who noted that they needed to call 911 for stroke symptoms increased from 80% to 87% in six months after project implementation. The results of this project demonstrate significant improvement in patients’ knowledge about the necessity of activation of emergency medical systems for stroke symptoms.

Keywords: emergency medical systems activation, evidence-based practice nursing, stroke education, video education

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Authors: H. Maachou, A. Chagnes, G. Cote

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The present work reports the properties of chitosan/hydroxyapatite (Cs/HA: 100/00, 70/30 and 30/70) composite microspheres obtained by emulsification processing route. The morphology of chitosane microspheres was observed by a scanning electron microscope (SEM) which shows an aggregate of spherical microspheres with a particle size, determined by optical microscope, ranged from 4 to 10 µm. Thereafter, a biomimetic approach was used to study the in vitro biomineralization of these composites. It concerns the composites immersion in simulated body fluid (SBF) for different times. The deposited calcium phosphate was studied using X-ray diffraction analysis (XRD), FTIR spectroscopy and ICP analysis of phosphorus. In fact, the mineral formed on Cs/HA microspheres was a mixture of carbonated HA and β-TCP as showed by FTIR peaks at 1419,5 and 871,8 cm-1 and XRD peak at 29,5°. This formation was induced by the presence of HA in chitosan microspheres. These results are confirmed by SEM micrographs which chow the Ca-P crystals growth in form of cauliflowers. So, these materials are of great interest for bone regeneration applications due to their ability to nucleate calcium phosphates in presence of simulated body fluid (SBF).

Keywords: hydroxyapatite, chitosan, microsphere, composite, bone regeneration

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Authors: Fernanda Gonçalves, Karla Alcântara, Vanessa Moura, Patrícia Nolasco, Jorge Kalil, Maristela Hernandez

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Introduction: Atherosclerosis is a chronic disease where two striking features are observed: retention of lipids and inflammation. Understanding the interaction between immune cells and lipoproteins involved in atherogenesis are urgent challenges, since cardiovascular diseases are the leading cause of death worldwide. Macrophages are critical to the development of atherosclerotic plaques and in the perpetuation of inflammation in these lesions. These cells are also directly involved in unstable plaque rupture. Recently different populations of macrophages are being identified in atherosclerotic lesions. Although the presence of M2 macrophages (macrophages activated by the alternative pathway, eg. The IL-4) has been identified, the function of these cells in atherosclerosis is not yet defined. M2 macrophages have a high endocytic capacity, they promote remodeling of tissues and to have anti-inflammatory activity. However, in atherosclerosis, especially unstable plaques, severe inflammatory reaction, accumulation of cellular debris and intense degradation of the tissue is observed. Thus, it is possible that the M2 macrophages have altered function (phenotype) in atherosclerosis. Objective: Our aim is to evaluate if the presence of oxidized LDL alters the phenotype and function of M2 macrophages in vitro. Methods: For this, we will evaluate whether the addition of lipoprotein in M2 macrophages differentiated in vitro with IL -4 induces 1) a reduction in the secretion of anti-inflammatory cytokines (CBA and ELISA), 2) secretion of inflammatory cytokines (CBA and ELISA), 3) expression of cell activation markers (Flow cytometry), 4) alteration in gene expression of molecules adhesion and extracellular matrix (Real-Time PCR) and 5) Matrix degradation (confocal microscopy). Results: In oxLDL stimulated M2 macrophages cultures we did not find any differences in the expression of the cell surface markers tested, including: HLA-DR, CD80, CD86, CD206, CD163 and CD36. Also, cultures stimulated with oxLDL had similar phagocytic capacity when compared to unstimulated cells. However, in the supernatant of these cultures an increase in the secretion of the pro-inflammatory cytokine IL-8 was detected. No significant changes where observed in IL-6, IL-10, IL-12 and IL-1b levels. The culture supernatant also induced massive extracellular matrix (produced by mouse embryo fibroblast) filaments degradation. When evaluating the expression of 84 extracellular matrix and adhesion molecules genes, we observed that the stimulation of oxLDL in M2 macrophages decreased 47% of the genes and increased the expression of only 3% of the genes. In particular we noted that oxLDL inhibit the expression of 60% of the genes constituents of extracellular matrix and collagen expressed by these cells, including fibronectin1 and collagen VI. We also observed a decrease in the expression of matrix protease inhibitors, such as TIMP 2. On the opposite, the matricellular protein thrombospondin had a 12 fold increase in gene expression. In the presence of native LDL 90% of the genes had no altered expression. Conclusion: M2 macrophages stimulated with oxLDL secrete the pro-inflammatory cytokine IL-8, have an altered extracellular matrix constituents gene expression, and promote the degradation of extracellular matrix. M2 macrophages may contribute to the perpetuation of inflammation in atherosclerosis and to plaque rupture.

Keywords: atherosclerosis, LDL, macrophages, m2

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Authors: Sandip Sharma, Jayesh Ruparelia

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The present work focuses on the comparative study of ozone based advanced oxidation processes (AOPs): O3, O3/UV and O3/UV/Persulfate for mineralization of synthetic wastewater containing Reactive Black5 (RB5) dye. The effect of various parameters: pH, ozone flow rate, initial concentration of dye and intensity of UV light was analyzed to access performance efficiency of AOPs. The performance of all the three AOPs was evaluated on the basis of decolorization, % TOC removal and ozone consumption. The highest mineralization rate of 86.83% was achieved for O3/UV/Persulfate followed by 71.53% and 66.82 % for O3/UV and O3 respectively. This is attributed to the fact that Persulfate ions (S2O82-) upon activation produce sulfate radical (SO4-●) which is very strong oxidant capable of degrading a wide variety of recalcitrant organic compounds, moreover to enhance the performance of Persulfate it is activated using UV irradiation. On increasing the intensity of UV irradiation from 11W to 66W, TOC removal efficiency is increased by 59.04%. Ozone based AOPs gives better mineralization on basic conditions, at pH 12 it gives 68.81%, 60.01% and 40.32% TOC removal for O3/UV/Persulfate, O3/UV and O3 process respectively. The result also reveals that decolorization of 98.95%, 95.17% and 94.71% was achieved by O3/UV/Persulfate, O3/UV and O3 process respectively. In addition to above, ozone consumption was also considerably decreased by 17% in case of O3/UV/Persulfate, as efficiency of process is enhanced by means of activation of persulfate through UV irradiation. Thus study reveals that mineralization follows: O3/UV/Persulfate> O3/UV> O3.

Keywords: AOP, mineralization, TOC, recalcitrant organic compounds

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Authors: Mingjun Xu, Man Pun Wan

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This study proposed an effective smoke control strategy for the large flat space with a low ceiling to achieve the requirement of perpetual tenability. For the large flat space with a low ceiling, the depth of the smoke reservoir is very shallow, and it is difficult to perpetually constrain the smoke within a limited space. A series of numerical tests were conducted to determine the smoke strategy. A zonal design i.e., the fire zone and two adjacent zones was proposed and validated to be effective in controlling smoke. Once a fire happens in a compartment space, the Engineered Smoke Control (ESC) system will be activated in three zones i.e., the fire zone, in which the fire happened, and two adjacent zones. The smoke can be perpetually constrained within the three smoke zones. To further improve the extraction efficiency, sequential activation of the ESC system within the 3 zones turned out to be more efficient than simultaneous activation. Additionally, the proposed zonal and sequential extraction design can reduce the mechanical extraction flow rate by up to 40.7 % as compared to the conventional method, which is much more economical than that of the conventional method.

Keywords: performance-based design, perpetual tenability, smoke control, fire plume

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Hairin Taha, Aditya Arya, M. A. Hapipah, A. M. Mustafa

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Plants have been an important source of medicine since ancient times. Pseuduvaria monticola is a rare montane forest species from the Annonaceae family. Traditionally, the plant was used to cure symptoms of fever, inflammation, stomach-ache and also to reduce the elevated levels of blood glucose. Scientifically, we have evaluated the antidiabetic potential of the Pseuduvaria monticola bark methanolic extract on certain in vitro cell based assays, followed by in vivo study. Results from in vitro models displayed PMm upregulated glucose uptake and insulin secretion in mouse pancreatic β-cells. In vivo study demonstrated the PMm down-regulated hyperglycaemia, oxidative stress and elevated levels of pro-inflammatory cytokines in type 2 diabetic rat models. Altogether, the study revealed that Pseuduvaria monticola might be used as a potential candidate for the management of type 2 diabetes and its related complications.

Keywords: type 2 diabetes, Pseuduvaria monticola, insulin secretion, glucose uptake

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Authors: Sina Gharevali

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Staphylococcus aureus is one of the most important factors for nosocomial infections and infections acquired in a hospital setting role as is. Drug-resistant bacteria methicillin, which in 1961 was reported in many parts of the world, Made the role as the last drug, vancomycin, in the treatment of infections caused by the Staphylococcus aureus chain be taken into consideration. The aim of this study was to evaluate the antimicrobial effects of copper nanoparticles and compared it with antibiotics on Staphylococcus aureus resistant to vancomycin in vitro and animal model. In this study, this test was performed, and the most effective antibiotic for vancomycin-resistant Staphylococcus aureus was determined by disk diffusion method. After various concentrations of copper nanoparticles and antibiotics were prepared and vancomycin resistant Staphylococcus aureus bacteria with serial dilution method for determining antibiotic ciprofloxacin. Minimum Inhibitory Concentration and Minimum Bactericidal Concentrationcopper nanoparticles was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. Then the broth dilution method for the antibiotic ciprofloxacin, nano-particles, and nano-particles of copper and copper-established antibiotic synergy MIC and MBC were obtained. MBC was obtained from the experimental animal model test method, and the results were compared. The results showed that copper nanoparticles compared with the antibiotic ciprofloxacin in vitro and animal model more effective in inhibiting the growth of Staphylococcus aureus resistant to vancomycin and ciprofloxacin and extent of the impact of the Synthetic effect of lower copper nanoparticles. Which can then be used to treat clinical research as a candidate.

Keywords: nanoparticles, copper, staphylococcus, aureus

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Authors: Pamita Awasthi, Kirna, Shilpa Dogra, Manu Vatsal, Ritu Barthwal

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Doxorubicin, also known as adriamycin, is an anthracycline class of drug used in cancer chemotherapy. It is used in the treatment of non-Hodgkin’s lymphoma, multiple myeloma, acute leukemias, breast cancer, lung cancer, endometrium cancer and ovary cancers. It functions via intercalating DNA and ultimately killing cancer cells. The major side effects of doxorubicin are hair loss, myelosuppression, nausea & vomiting, oesophagitis, diarrhoea, heart damage and liver dysfunction. The minor modifications in the structure of compound exhibit large variation in the biological activity, has prompted us to carry out the synthesis of sulfonamide derivatives. Sulfonamide is an important feature with broad spectrum of biological activity such as antiviral, antifungal, diuretics, anti-inflammatory, antibacterial and anticancer activities. Structure of the synthesized compound N-(1-methyl-2-oxo-2-N-methyl anilino-ethyl)benzene sulfonamide confirmed by proton nuclear magnetic resonance (1H NMR),13C NMR, Mass and FTIR spectroscopic tools to assure the position of all protons and hence stereochemistry of the molecule. Further we have reported the binding potential of synthesized sulfonamide analogues in comparison to doxorubicin drug using Auto Dock 4.2 software. Computational binding energy (B.E.) and inhibitory constant (Ki) has been evaluated for the synthesized compound in comparison of doxorubicin against Poly (dA-dT).Poly (dA-dT) and Poly (dG-dC).Poly (dG-dC) sequences. The in vitro cytotoxic study against human breast cancer cell lines confirms the better anticancer activity of the synthesized compound over currently in use anticancer drug doxorubicin. The IC50 value of the synthesized compound is 7.12 µM where as for doxorubicin is 7.2 µ.

Keywords: Doxorubicin, auto dock, in silco, in vitro

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Authors: Olumayede Emmanuel Gbenga, Adeniyi Azeez Adebayo

Abstract:

Carbonyls are the first-generation products from tropospheric degradation reactions of volatile organic compounds (VOCs). This computational study examined the mechanism of removal of carbonyls from the atmosphere via hydroxyl radical. The kinetics of the reactions were computed from the activation energy (using enthalpy (ΔH**) and Gibbs free energy (ΔG**). The minimum energy path (MEP) analysis reveals that in all the molecules, the products have more stable energy than the reactants, which implies that the forward reaction is more thermodynamically favorable. The hydrogen abstraction of the aromatic aldehyde, especially without methyl substituents, is more kinetically favorable compared with the other aldehydes in the order of aromatic (without methyl or meta methyl) > alkene (short chain) > diene > long-chain aldehydes. The activation energy is much lower for the forward reaction than the backward, indicating that the forward reactions are more kinetically stable than their backward reaction. In terms of thermodynamic stability, the aromatic compounds are found to be less favorable in comparison to the aliphatic. The study concludes that the chemistry of the carbonyl bond of the aldehyde changed significantly from the reactants to the products.

Keywords: atmospheric carbonyls, oxidation, mechanism, kinetic, thermodynamic

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Authors: Sunil Kumar, Sandip Dhole, Deepak Salunke, Chung-ming Sun

Abstract:

Heterocycles bearing nitrogen, oxygen, and selenium are present in innumerable biologically active compounds. For instance, coumarin containing dicoumarol acts as naturally occurring anticoagulant. 2-Acylamido selenazole works as Store-Operated Calcium (SOC) channel regulator. Therefore, due to biologically significance of selenazole and coumarin and our quest to develop efficient methodologies for the synthesis of complex heterocycles, the trisubstituted angular isocoumarinoselenazole synthesis was proposed and achieved by starting from nitrobenzoic acid derivative, available commercially. Synthetic procedure involves three steps: i) the construction of 2-aminobenzoselenazoles, ii) their regioselective N-alkylation at position-2 and iii) alkyne insertion via Ru catalyzed C-H activation. Transition metal free synthesis of benzoselenazoles was successfully brought about by the addition/elimination reaction via intramolecular C-Se bond formation. In the next step, N-alkylation of selenazole furnished two regioisomers. Both the isomers exhibited different reactivity towards [4+2] alkyne annulation reaction. The fusion of α-pyrone ring on the benzo[1,3-d]selenazole skeleton was achieved via Ru(II)-catalyzed C-H activation and alkyne insertion. As evident from mechanism, the selenazole 'N' plays an important role for the experiential selectivity.

Keywords: alkylation, alkyne insertion, coumarin, selenazole

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Authors: Eunseop Yeom

Abstract:

Platelets can be activated by the surrounding blood flows where a blood vessel is narrowed as a result of atherosclerosis. Numerous studies have been conducted to identify the relation between platelets activation and thrombus formation. To measure platelet adhesion, this study proposes an image analysis technique. Blood samples are delivered in the microfluidic channel, and then platelets are activated by a stenotic micro-channel with 90% severity. By applying proposed correlation mapping, which visualizes decorrelation of the streaming blood flow, the area of adhered platelets (APlatelet) was estimated without labeling platelets. In order to evaluate the performance of correlation mapping on the detection of platelet adhesion, the effect of tile size was investigated by calculating 2D correlation coefficients with binary images obtained by manual labeling and the correlation mapping method with different sizes of the square tile ranging from 3 to 50 pixels. The maximum 2D correlation coefficient is observed with the optimum tile size of 5×5 pixels. As the area of the platelet adhesion increases, the platelets plug the channel and there is only a small amount of blood flows. This image analysis could provide new insights for better understanding of the interactions between platelet aggregation and blood flows in various physiological conditions.

Keywords: platelet activation, correlation coefficient, image analysis, shear rate

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Authors: Davender Kumar

Abstract:

Aim: It is important for the orthodontist to be familiar with the sliding resistance (SR) generated by the ligation method used during the space closure phase with sliding mechanics. To determine new, experimental non-conventional (slide) ligature demonstrates less friction in vitro when compared other ligatures on the market. Methods: Experimental in vitro were carried out to test the performance of the low-friction system with regard to assess the forces released by different bracket–ligature systems with bonded in iron plate mounted on an Instron machine. Results: The outcomes of experimental testing showed that the combination of the low-friction ligatures with the super elastic nickel-titanium and SS wires produced a significantly smaller amount of binding at the bracket/arch wire/ligature unit when compared to conventional elastomeric ligatures. Conclusion: The biomechanical consequences of the use of low-friction ligatures were shorter duration of orthodontic treatment during the levelling and aligning phase, concurrent dentoalveolar expansion of the dental arch, and the possibility of using biologically adequate orthodontic forces.

Keywords: archwire, bracket, friction, ligation

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Authors: Nusaiba Al-Nemrawi, Belal Al-Husein

Abstract:

Methotrexate (MTX) is a stoichiometric inhibitor of dihydrofolate reductase, which is essential for DNA synthesis. MTX is a chemotherapeutic agent used for treating many types of cancer cells. However, cells’ resistant to MTX is very common and its pharmacokinetic behavior is highly problematic. of MTX within tumor cells, we propose encapsulation of antitumor drugs in nanoparticulated systems. Chitosan (CS) is a naturally occurring polymer that is biocompatibe, biodegradable, non-toxic, cationic and bioadhesive. CS nanoparticles (CS-NPs) have been used as drug carrier for targeted delivery. Titanium dioxide (TiO2), a natural mineral oxide, which is used in biomaterials due to its high stability and antimicrobial and anticorrosive properties. TiO2 showed a potential as a tumor suppressor. In this study a new formulation of MTX loaded in CS NPs (CS-MTX NPs) and coated with Titanium oxide (TiO2) was prepared. The mean particle size, zeta potential, polydispersity index were measured. The interaction between CS NPs and TiO2 NPs was confirmed using FTIR and XRD. CS-MTX NPs was studied in vitro using the tumor cell line MCF-7 (human breast cancer). The results showed that CS-MTX has a size around 169 nm and as they were coated with TiO2, the size ranged between and depending on the ratio of CS-MTX to TiO2 ratio used in the preparation. All NPs (uncoated and coated carried positive charges and were monodispersed. The entrapment efficacy was around 65%. Both FTIR and XRD proved that TiO2 interacted with CS-MTX NPs. The drug invitro release was controlled and sustained over days. Finally, the studied in vitro using the tumor cell line MCF-7 suggested that combining nanomaterials with anticancer drugs CS-MTX NPs may be more effective than free MTX for cancer treatment. In conclusion, the combination of CS-MTX NPs and TiO2 NPs showed excellent time-dependent in vitro antitumor behavior, therefore, can be employed as a promising anticancer agent to attain efficient results towards MCF-7 cells.

Keywords: Methotrexate, Titanium dioxide, Chitosan nanoparticles, cancer

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Authors: Eman Alzahrani

Abstract:

Date palm trees are abundant and cheap natural resources in Saudi Arabia. In this study, an activated carbon was prepared from palm trunks by chemical processes. The chemical activation was performed by impregnation of the raw materials after grinding with H3PO4 solution (63%), followed by placing of the sample solution on a muffle furnace at 400ºC for 30 min, and then at 800ºC for 10 min. The morphology of the fabricated material was checked using scanning electron microscopy that showed the rough surfaces on the carbon samples. The use of fabricated activated carbon for removal of eosin dye from aqueous solutions at different contact time, initial dye concentration, pH and adsorbent doses was investigated. The experimental results show that the adsorption process attains equilibrium within 20 min. The adsorption isotherm equilibrium was studied by means of the Langmuir and Freundlich isotherms, and it was found that the data fit the Langmuir isotherm equation with maximum monolayer adsorption capacity of 126.58 mg g-1. The results indicated that the home made activated carbon prepared from palm trunks has the ability to remove eosin dye from aqueous solution and it will be a promising adsorbent for the removal of harmful dyes from waste water.

Keywords: activated carbon, date palm trunks, H3PO4 activation, adsorption, dye removal, eosin dye, isotherm

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Authors: Nisha Nandlal Sharma, Julaluk Carmai, Saiprasit Koetniyom, Bernd Markert

Abstract:

Whiplash Injuries are mostly experienced in car accidents. Symptoms of whiplash are commonly reported in studies, neck pain and headaches are two most common symptoms observed. The whiplash Injury mechanism is poorly understood. In present study, hybrid neck muscle model were developed with a combination of solid tetrahedral elements and 1D beam elements. Solid tetrahedral elements represents passive part of the muscle whereas, 1D beam elements represents active part. To simulate the active behavior of the muscle, Hill-type muscle model was applied to beam elements. To simulate non-linear passive properties of muscle, solid elements were modeled with rubber/foam material model. Some important muscles were then inserted into THUMS (Total Human Model for Safety) THUMS was given a boundary conditions similar to experimental tests. The model was exposed to 4g and 7g rear impacts as these load impacts are close to low speed impacts causing whiplash. The effect of muscle activation level on occupant kinematics during whiplash was analyzed.

Keywords: finite element model, muscle activation, THUMS, whiplash injury mechanism

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Authors: Prakriti Diwan, S. Saraf

Abstract:

The present study involves preparation and evaluation of microparticles of diclofenac sodium. The microparticles were prepared by the emulsion solvent evaporation techniques using ethylcellulose polymer. Four different batches of microspheres were prepared by varying the concentration of polymer from 50% to 80% w/w. The microspheres were characterized for drug content, percentage yield and encapsulation efficiency, particle size analysis and surface morphology. Microsphere prepared with high drug content produces higher percentage yield and encapsulation efficiency values. It was observed the increase in concentration of the polymer, increases the mean particle size of the microspheres. The effect of polymer concentration on the in vitro release of diclofenac from the microspheres was also studied. The production microparticles yield showed 98.74%, mean particle size 956.32µm and loading efficiency 97.15%. The results were found that microparticles prepared had slower release than microparticles (p>0.05). Therefore, it may be concluded that drug loaded microparticles are suitable delivery systems for diclofenac sodium.

Keywords: diclofenac sodium, emulsion solvent evaporation, ethylcellulose, microparticles

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Authors: Adil Elrayah, Jie Weng, Esra Suliman

Abstract:

The mineral in human bone is not pure stoichiometric calcium phosphate (Ca/P) as it is partially substituted by in organic elements. In this study, the copper ions (Cu²⁺) substituted hydroxyapatite (CuHA) powder has been synthesized by the co-precipitation method. The CuHA powder has been used to fabricate CuHA fiber scaffolds by sol-gel process and the following sinter process. The resulted CuHA fibers have slightly different microstructure (i.e. porosity) compared to HA fiber scaffold, which is denser. The mechanical properties test was used to evaluate CuHA, and the results showed decreases in both compression strength and hardness tests. Moreover, the in vitro used endothelial cells to evaluate the angiogenesis of CuHA. The result illustrated that the viability of endothelial cell on CuHA fiber scaffold surfaces tends to antigenic behavior. The results obtained with CuHA scaffold give this material benefit in biological applications such as antimicrobial, antitumor, antigens, compacts, filling cavities of the tooth and for the deposition of metal implants anti-tumor, anti-cancer, bone filler, and scaffold.

Keywords: fiber scaffold, copper ions, hydroxyapatite, in vitro, mechanical property

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