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      • KCI등재

        An Insight Into the Physico-Mechanical Signatures of Silylated Graphene Oxide in Poly(ethylene methyl acrylate) Copolymeric Thermoplastic Matrix

        Sayan Ganguly,Subhadip Mondal,Poushali Das,Poushali Bhawal,Tushar Kanti Das,Sabyasachi Ghosh,Sanjay Remanan,Narayan Chandra Das 한국고분자학회 2019 Macromolecular Research Vol.27 No.3

        Dispersion of graphene as nano-building block in polymer matrix is challenging for developing high strength polymer nanocomposites. Tuning of surface polarity can be an effective pathway to resolve this issue of dispersion. Besides this, the polymer matrix (Ethylene methyl acrylate or EMA) has been chosen here judicially due to its polar-nonpolar alternating copolymeric segments which indirectly facilitated dispersion of nanofillers. Herein, graphene oxide has been lyophilically modified by virtue of surface grafting phenomenon with the help of di-halo substituted silane. The most surprising outcome which has been nurtured is their superior dispersion, improvement in physico-mechanical features, and transparency without affecting the inherent compliance of pristine polymer. The transmission electron microscopic image of silane functionalized graphene oxide (GOF) is showing surface roughness which has immense effect of physisorption and mechanical anchoring of polymer chains over GOF nano-sheets. Such physical interaction has enough impact on mechanical properties which has been discussed here. Moreover, the deterioration of transparency was not so much affected after loading of GOF filler. The filler distribution also has been confirmed in the light of small angle X-ray scattering (SAXS) study. Thermal treatment has been conducted for composites which accounted high thermal stability comparatively to pristine polymer.

      • Expansion of human mesenchymal stem cells using the wave bioreactor system with pulsed electromagnetic field stimulation for enhanced cell culture performance

        ( Sayan Deb Dutta ),( Keya Ganguly ),( Dinesh Patel ),( Tejal Patel ),( Ki-taek Lim ) 한국농업기계학회 2021 한국농업기계학회 학술발표논문집 Vol.26 No.2

        Most bone tissue-engineering models fail to demonstrate the complex cellular functions of living bone; therefore, most translational studies on bone tissue are performed in live models. To reduce the need for live models, we developed a stimulated micro-chip model for monitoring protein secretion during osteogenesis using human mesenchymal stem cells (hBMSCs). We established a bone micro-chip system for monitoring the in vitro differentiation and sensing the secreted proteins of hMSCs under a sinusoidal electromagnetic field (SEMF), which ameliorates bone healing in a biomimetic natural bone matrix. A 3V-1Hz SEMF biophysically stimulated osteogenesis by activating ERK-1/2 and promote phosphorylation of p38 MAPK kinases. Exposure to a 3V-1Hz SEMF upregulated the expression of osteogenesis-related genes, and enhanced the expression of key osteoregulatory proteins. We identified 23 proteins that were differentially expressed in stimulated hBMSC secretomes, or were absent in the control groups. Our on-chip stimulation technology is easy to use, versatile, and non-disruptive, and should have diverse applications in regenerative medicine and cell-based therapies.

      • 3D printing of a conductive polypyrrole-grafted gelatin methacrylate (GelMA)-based hydrogels for continuous microcurrent stimulation of human mesenchymal stem cells

        ( Sayan Deb Dutta ),( Keya Ganguly ),( Tejal Patel ),( Ki-taek Lim ) 한국농업기계학회 2021 한국농업기계학회 학술발표논문집 Vol.26 No.2

        Electrical stimulation has been shown to ameliorate bone healing for a long time. This study developed a hybrid and 3D printable conductive methacrylated gelatin-polypyrrole (GelMA-PPy)-based photocurable and self-healing hydrogel inks for continuous microcurrent stimulation. For this, a custom-made electrical stimulation device (DC stimulation) was used to evaluate the osteogenic differentiation of human bone mesenchymal stem cells (hMSCs). The pyrrole was chemically grafted onto the surface of GelMA via a one-step conjugation reaction with ammonium persulfate (APS) and cross-linked with iron (III) chloride. The fabricated hydrogel was characterized by proton nuclear magnetic resonance (1H-NMR), scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FT-IR) to analyze the grafting and chemical interaction. The as-prepared hydrogel exhibited higher mechanical and swelling properties compared to pure GelMA scaffolds. The 3D printed GelMA-PPy hydrogel showed improved cell viability (~12.6%) when exposed to 500 mV/day current, compared to pure GelMA after 7 and 14 days of cell culture. Interestingly,16.0~20.0-fold higher expression of osteogenic genes and protein markers (Runx2, ALP, OCN, and OPN) were observed in the GelMA-PPy treated groups than control suggest that pyrrole incorporation into GelMA matrix significantly improved the conductivity and osteogenic differentiation of hMSCs. Therefore, this study shows that the hMSCs react differentially to low-voltage DC electrical stimulation in the presence of GelMA-PPy scaffolds, which could be used as an ideal material for electrical stimulation for tissue engineering.

      • Expansion of human mesenchymal stem cells through wave motion bioreactor system with pulsed electromagnetic field stimulation for enhanced cell culture and real-time glucose monitoring

        ( Sayan Deb Dutta ),( Keya Ganguly ),( Tejal Patel ),( Ki-taek Lim ) 한국농업기계학회 2021 한국농업기계학회 학술발표논문집 Vol.26 No.2

        Mesenchymal stem cells (hMSCs) are considered as one of the promising approaches for treating several diseases in cell-based therapeutics. The current strategies for hMSCs expansion include the in vitro static culture system yielding a low number of cells. Therefore, a scalable, dynamic bioreactor-based culture system is needed to produce an adequate number of cells for clinical applications. In this study, a new strategy for hMSCs expansion is employed with a modified wave-motion bioreactor system coupled with continuous electromagnetic field (EMF) stimulation. For this purpose, hMSCs were tested in various EMF exposure (1V, 5V, and 10V-1Hz) to find the maximum viability for cell expansion. Interestingly, the hMSCs tend to form aggregate-like structures during the wave motion culture. We show that a 10V-1Hz EMF exposure (magnetic field: 5.82 G) during wave motion culture (35 rpm) induces the viability of hMSCs up to 15% than static culture as evaluated by WST-8 assay. Moreover, the long-term cell culture of hMSCs in wave bioreactor significantly enhanced the expression of osteogenic transcription factors, such as Runx2 (6.8-8.0 fold), ALP (8.0 fold), BSP (5-6.0 fold), and OCN (8-10.0 fold) compared to the static culture (28% higher as of control). We also found that the average glucose consumption was significantly higher compared to static culture as predicted by Six biosensor B.LV5. In the initial phase of cell culture, the average detectable level of glucose was 8.35±0.34 mM/μL and lactate was 0.63±0.05 mM/μL. However, after 7 days of dynamic culture, the available glucose in the media was estimated about 2.57±0.32 mM/μL and lactate was 7.67±0.26 mM/μL which was significantly higher than the static control group. The RNA-sequencing results further indicate the upregulation of major osteogenic/mechanically stimulated signaling pathways during wave motion culture of hMSCs. These results suggested that the use of 10V-1Hz (5.82 G) EMF is non-toxic for hMSCs, and the modification of wave bioreactor with externally controlled EMF has good potential for hMSCs proliferation and expansion.

      • Electromagnetic field-stimulated microreactor system with biosensing potential of stem cell-secreted proteins

        ( Sayan Deb Dutta ),( Tusan Park ),( Keya Ganguly ),( Ki-taek Lim ) 한국농업기계학회 2021 한국농업기계학회 학술발표논문집 Vol.26 No.2

        Most bone tissue-engineering models fail to demonstrate the complex cellular functions of living bone; therefore, most translational studies on bone tissue are performed in live models. To reduce the need for live models, we developed a stimulated micro-chip model for monitoring protein secretion during osteogenesis using human mesenchymal stem cells (hBMSCs). We established a bone micro-chip system for monitoring the in vitro differentiation and sensing the secreted proteins of hMSCs under a sinusoidal electromagnetic field (SEMF), which ameliorates bone healing in a biomimetic natural bone matrix. A 3V-1Hz SEMF biophysically stimulated osteogenesis by activating ERK-1/2 and promote phosphorylation of p38 MAPK kinases. Exposure to a 3V-1Hz SEMF upregulated the expression of osteogenesis-related genes, and enhanced the expression of key osteoregulatory proteins. hBMSCs undergoing osteoblastic differentiation are responsive to a well-defined SEMF stimulation at 3 V and 1 Hz, with an amplitude of 0.1 mT, for 20 minutes once a day. SEMF-treated plates contained the highest number of viable cells (>11%) compared to the other groups. Constant treatment with higher voltage, amplitude, or for a longer duration is generally counterproductive, suggesting that these parameters are optimized for this cell type. Our findings suggest that low voltage-frequency SEMF-guided osteoblast differentiation is not only triggered by the exposure time or dosage, but is also affected by various physicochemical factors. The activation of ERK1/2 and phosphorylation of p38 factors regulates the osteogenic differentiation during SEMF stimulation. Moreover, the bioinformatics analysis revealed the activation of various genes that are differentially expression only during SEMF stimulation but not in unstimulated condition. Notably, the gene expression of ALP (1.3 fold) and OCN (5.2 fold) was significantly higher (*p<0.05, **p<0.01) in cells receiving 3 V-1 Hz (0.1 mT, 20 min/day) stimulation compared to in the control. Significant increases in the expression of osteogenic gene markers were also observed for Runx2, OSX, BSP, OPN, and COL1, compared to in the unstimulated cells (*p<0.05). Six secretome proteins were upregulated in stimulated hBMSCs compared to in unstimulated hBMSCs. Overall, 23 proteins were either upregulated or downregulated by SEMF stimulation. These proteins are predicted to be secreted or included in secretory vesicles according to annotations in the Uniport database. Out of the upregulated proteins in SEMF-stimulated hBMSCs, 36% or 50% are involved in the 'immune response' or 'extracellular matrix function,' respectively. The unprecedented efficacy of our low-voltage-frequency SEMF exposure protocol for achieving hBMSC osteogenesis has broad clinical and practical implications, and could form the basis of SEMF-based therapeutic strategies for stem cell-based bone tissue regeneration. Our on-chip stimulation technology is easy to use, versatile, and non-disruptive, and should have diverse applications in regenerative medicine and cell-based therapies.

      • Electromagnetic field-assisted cell-laden 3D printed poloxamer-407 hydrogel for enhanced osteogenesis

        ( Sayan Deb Dutta ),( Keya Ganguly ),( Tejal Patel ),( Ki-taek Lim ) 한국농업기계학회 2021 한국농업기계학회 학술발표논문집 Vol.26 No.2

        3D bioprinted hydrogel has gained enormous attention, especially in tissue engineering, owing to its attractive structure and excellent biocompatibility. In this study, we demonstrated that 3D bioprinted cell-laden ‘thermoresponsive’ poloxamer-407 (P407) gels have the potential to stimulate osteogenic differentiation of apical papilla stem cells (SCAPs) under the influence of low voltage- frequency 1V-1Hz (0.14 mT), 5V-1Hz (0.62 mT), and 10V-1Hz (1.21 mT) electromagnetic fields (EMFs). Their exposure time was 5 min/day, 10 min/day, 20 min/day, and 30 min/day for each hydrogel group, respectively. The developed hydrogel exhibited higher mechanical strength as well as good printability, showing high-quality micro-architecture. Moreover, the as-printed hydrogels (5 mm × 5 mm) were loaded with plasminogen activator inhibitor-1 (PAI-1) for testing the combined effect of PAI-1 and EMFs on SCAP differentiation. Interestingly, the 3D hydrogels showed improved viability and differentiation of SCAPs under EMFs' influence as examined by live/dead assay and alizarin Red-S staining, respectively. Our results demonstrated that DSPP and DMP-1 markers' expression significantly increased (3.8-fold) in 5V (0.62 mT) EMFs treatment. A similar fashion was also observed in ALP and Col-1(2.80-fold), comparable to the control groups. Therefore, the higher expression of theses gene markers (DSPP, DMP-1 ALP, and Col-1) indicated their better osteogenic efficiency of P407-encapsulated SCAPs in the presence of EMFs. The results confirmed that P407 hydrogels are non-toxic for encapsulation of SCAPs, yielding high cell viability and accelerate the cell migration potential. The 3D hydrogels with PAI-1 exhibited high mRNA expression levels for osteogenic/odontogenic gene markers (ALP, Col-1, DSPP, and DMP-1) vis-à-vis control after 14 days of in vitro culture. Our findings suggest that 3D bioprinted P407 hydrogels are biocompatible for SCAP encapsulation, and the applied low voltage-frequency EMFs could effectively improve dental tissue regeneration, particularly for oral applications.

      • Fluid shear stress-induced osteogenic differentiation of human mesenchymal stem cells on a 3D printed nanohydroxyapatite-GelMA hydrogel by a custom-designed six well perfusion chamber system

        ( Keya Ganguly ),( Sayan Deb Dutta ),( Tejal Patel ),( Ki-taek Lim ) 한국농업기계학회 2021 한국농업기계학회 학술발표논문집 Vol.26 No.2

        Bone tissue engineering has emerged as an excellent alternative to the traditional treatment strategies of bone injuries. However, current in-vitro systems do not fully mimic the biological environment of the bone defects. Hence, the design of bioreactor system is worth investing to allow monitoring of the de-novo bone formation in vitro. Herein, we have evaluated the osteogenic differentiation of human bone marrow derived mesenchymal stem cells seeded on to 3D printed nanohydroxyapatite-GelMA (nHAp-GelMA) hydrogel under the different flow rates (0.1 mL/ min, 0.25 mL/ min, and 0.5 mL/ min) in a custom designed perfusion type bioreactor. The mechanical property of the as-fabricated hydrogel increased as the concentration of hydroxyapatite increased from 0.5 to 2%, respectively. Moreover, the incorporation of nHAp into GelMA matrix also improved the swelling efficiency and mineralization potential in vitro. Our results indicated a higher osteogenic differentiation in the hBMSCs under the fluid shear stress as compared to the static cultures. A 10~15% increase in the cell viability was observed under the varying fluid shear stress as determined by the WST-8 assay and the Live-Dead cell viability assay. Moreover, an enhancement in the overall osteogenic differentiation markers in the hBMSCs under the fluid shear stress (as of control) was evident by the qRT-PCR analysis of the osteogenic marker genes including ALP (7.0~9.5-fold), Runx2 (16.5-fold), OSX (6.5-fold), Col1 (18.0-fold), OPN (10.8-fold), and BSP (12.5-fold). The upregulation of the respective proteins was also supported by the transcriptome analysis. Also, applied shear stress induced the secretion of several growth factors and cytokines. Taken together, our study indicated fluid-shear stress by the custom-designed perfusion chamber induces hBMSC differentiation and in vitro bone regeneration.

      • Pulsatile pressure mechanical stimulation on 3D printed gelatin methacrylate (GelMA)/ cellulose nanocrystals hybrid hydrogel promotes osteogenic differentiation of human mesenchymal stem cells

        ( Keya Ganguly ),( Sayan Deb Dutta ),( Dinesh Patel ),( Tejal Patel ),( Ki-taek Lim ) 한국농업기계학회 2021 한국농업기계학회 학술발표논문집 Vol.26 No.2

        The rapid infiltration and osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) on 3D printed osteogenic scaffolds represents a promising strategy to promote bone remodeling. However, the effect of constant wear and tear resulting from physical activities on mechanical integrity and functionality of tissue engineering constructs is crucial to determine before scaffold implantation. Therefore, we have focused on constructing an indigenous pulsatile bioreactor system creating artificial pressure loading microenvironment on GelMA/cellulose nanocrystals 3D-printed hydrogel to analyze the osteogenic differentiation of hBMSCs under a constant pulsatile pressure stimulation. The osteogenic differentiation potential of the hBMSCs seeded onto the fabricated scaffold was monitored after 7 and 14 days of cell culture under a pulsatile regiment of 10 ± 0.8 kPa pressure stimulation. Concurrently, hBMSCs maintained under static condition was taken as the control set. Our results indicated a 14.8% increase in the hBMSCs viability analyzed through the WST-8 assay and Live-Dead assay. Besides, increased adherence efficiency was evident by the morphometric analysis of the actin cytoskeletal arrangements and the expression of focal adhesion protein (Paxillin) determined by the immunocytochemistry analysis. Moreover, the qRT-PCR results after 7 days of cell culture indicated a higher expression of the early osteogenic marker genes, including ALP (7-fold), Runx2 (10.0-fold), OSX (5.8-fold), and mechanosensitive marker genes YAP/TAZ (4.2-fold), confirming the massive onset of early osteogenic differentiation in the hBMSCs. Similarly, the expression of the late osteogenic marker genes, including COL1 (8.0-fold), OPN (15.0-fold), and BSP (12.0-fold), was noted to be higher in cells cultured under pressure stimulation after 14 days of cell culture. Besides, the transcriptome analysis revealed the regulation of mechanosensitive genes during pressure stimulation than static group. The pulsatile pressure mechanical stimulation can be used to predetermine the mechanical potential of bone constructs and rapid osteogenic differentiation of hBMSCs.

      • KCI등재

        Fabrication of Reduced Graphene Oxide/Silver Nanoparticles Decorated Conductive Cotton Fabric for High Performing Electromagnetic Interference Shielding and Antibacterial Application

        Sabyasachi Ghosh,Sayan Ganguly,Poushali Das,Tushar Kanti Das,Madhuparna Bose,Nikhil K. Singha,Amit Kumar Das,Narayan Ch. Das 한국섬유공학회 2019 Fibers and polymers Vol.20 No.6

        Conductive filler loading in the polymer matrix is a common practice to transform insulative polymers toconducting composites. In case of textiles, the highly promising approach has been coined by virtue of fabricating withconductive adhesive homogeneous coating. The present fabrication approach has been developed by two-stage wet mixingtechnique including synthesis of silver nanoparticles decorated graphene sheets (rGO/Ag), followed by the preparation ofconducting coating by non-ionic polymer adhesive. The novelty lies in the choice of conductive material and coating strategyto make lightweight and flexible smart electronic fabric. In order to protect the radiation pollution from the immense use ofelectronic devices and gadgets, the coated textiles can be an excellent replacement of other commercially available polymercoatings. The electromagnetic interference (EMI) shielding effectiveness of the prepared coated textile was 27.36 dB in the Xband (8.2-12.4 GHz). Besides this it is worth mentioning that our developed coated fabric was enough conductive to light upa series of 57 LEDs with high intensity. Last but not the least this work also reconnoitres bactericidal feature against E. coli.

      • KCI등재

        Carbon Nanostructures Based Mechanically Robust Conducting Cotton Fabric for Improved Electromagnetic Interference Shielding

        Sabyasachi Ghosh,Subhadip Mondal,Sayan Ganguly,Sanjay Remanan,Nikhil Singha,Narayan Ch. Das 한국섬유공학회 2018 Fibers and polymers Vol.19 No.5

        Herein, an intelligent cotton fabric was fabricated using a non-ionic surfactant based macro structured carbonaceous coating through the ‘knife-over-roll’ technique. The developed novel fabric was tested as flexible, mechanically robust with prolonged chemical/moisture resistance. Various characterization techniques were thoroughly used to analyze the fabric. The as-prepared fabric shows an outstanding electromagnetic interference (EMI) shielding efficiency (SE) of about 21.5 dB even at the lowest possible coating thickness (0.20 mm) where the highest EMI SE of 30.8 dB is obtained at only 0.30 mm coating thickness over the X-band frequency range (8.2-12.4 GHz), possibly due to the threedimensionally interconnected network structure of conducting carbon particles. The micro-computed tomography disclosed the porous architecture and “void-filler” arrangement within the fabrics. For the betterment of serviceability and practicability of the coated fabric, the water tolerance and contact angle studies were conducted. The relatively high contact angle than pure cotton fabric, and excellent water resistance after coating ensure improved endurance for external or industrial uses. Therefore, this proof-of-construct manifests commercialization of the developed fabric for multipurpose applications in a facile, less-hazardous and economical way.

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