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1.
Int J Biol Macromol ; 116: 774-785, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29777811

ABSTRACT

Fabrication of mechanically stable, biocompatible bilayered polymeric scaffold consisting of chitosan(CS)/polycaprolactone(PCL) and hyaluronic acid(HA) using less toxic solvent system is presented in this study. Electrospinning technique to make the scaffold was used followed by morphological, physiochemical and mechanical characterizations. Average fiber diameter of CS/PCL-HA bilayered scaffold was found 362.2 ±â€¯236 nm which is in the range of collagen fiber found in the extracellular matrices. Enhanced swelling, degradation, hydrophilicity and water vapour transmission rate were found for the bilayered scaffold compared to that of the PCL and CS-PCL scaffolds. Antimicrobial property evaluation revealed reduction in bacterial adhesion on bilayered scaffolds. Invitro studies with vero cells [kidney epithelial cell, extracted from African Green Monkey (Chlorocebus sp.)] confirm enhanced proliferation, growth and migration of vero cell on the bilayered CS/PCL-HA scaffold to that of PCL and CS/PCL scaffolds. Novelty of this study includes the use of HA for mechanically stabilized scaffold with acceptable biological properties for wound healing applications.


Subject(s)
Cell Proliferation/drug effects , Chitosan , Collagen , Extracellular Matrix/chemistry , Polyesters , Wound Healing/drug effects , Animals , Chitosan/chemistry , Chitosan/pharmacology , Chlorocebus aethiops , Collagen/chemistry , Collagen/pharmacology , Polyesters/chemistry , Polyesters/pharmacology , Vero Cells
2.
Mol Carcinog ; 54(12): 1636-55, 2015 Dec.
Article in English | MEDLINE | ID: mdl-25358602

ABSTRACT

Reactive oxygen species (ROS), the key mediators of cellular oxidative stress and redox dysregulation involved in cancer initiation and progression, have recently emerged as promising targets for anticancer drug discovery. Continuous free radical assault upsets homeostasis in cellular redox system and regulates the associated signaling pathways to mediate stress-induced cell death. This study investigates the dose-specific pro-oxidative behavior of a bacterial fucose polysaccharide, which attenuated proliferation of different cancer cells. In the fermentation process, Bacillus megaterium RB-05 [GenBank Accession Number HM371417] was found to biosynthesize a polysaccharide with low-fucose content (4.9%), which conferred the maximum anti-proliferative activity (750 µg/mL) against human lung cancer epithelial cells (A549) during preliminary screening. Structural elucidation and morphological characterization of the duly purified polysaccharide was done using HPLC, GC-MS, (1)H/(13)C NMR, and microscopy. The polysaccharide exhibited concentration- and time-dependent anti-proliferative effects against A549 cells by inducing intracellular ROS level and regulating the mitochondrial membrane-permeability following the apoptotic pathway. This process encompasses activation of caspase-8/9/3/7, increase in the ratio of Bax/Bcl2 ratio, translocation of Bcl2-associated X protein (Bax) and cytochrome c, decrease in expression of anti-apoptotic members of Bcl2 family, and phosphorylation of mitogen activated protein kinases (MAPKs). Apoptosis was attenuated upon pretreatment with specific caspase-inhibitors. Simultaneously, during apoptosis, the ROS-mediated stress as well as activated MAPKs triggered nuclear translocation of transcription factors like nuclear factor (erythroid-derived)-like 2 (Nrf2) and promoted further transcription of downstream cytoprotective genes, which somehow perturbed the chemotherapeutic efficacy of the polysaccharide, although using CuPP, a chemical inhibitor of HO-1, apoptosis increased significantly (P < 0.05).


Subject(s)
Apoptosis/drug effects , Intracellular Signaling Peptides and Proteins/metabolism , Lung Neoplasms/metabolism , Mitochondria/drug effects , Mitogen-Activated Protein Kinase Kinases/metabolism , NF-E2-Related Factor 2/metabolism , Polysaccharides, Bacterial/pharmacology , Apoptosis Regulatory Proteins/metabolism , Carcinoma/metabolism , Cell Line, Tumor , Epithelial Cells/metabolism , Fucose/pharmacology , HeLa Cells , Homeostasis/drug effects , Humans , Kelch-Like ECH-Associated Protein 1 , MCF-7 Cells , Mitochondria/metabolism , Reactive Oxygen Species/metabolism , Signal Transduction/drug effects
3.
Bioresour Technol ; 102(11): 6629-32, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21507629

ABSTRACT

Bacillus megaterium RB-05 was grown on glucose and on "tossa-daisee" (Corchorus olitorius)-derived jute, and production and composition of extracellular polysaccharide (EPS) were monitored. An EPS yield of 0.065 ± 0.013 and of 0.297 g ± 0.054 g(-1) substrate after 72 h was obtained for glucose and jute, respectively. EPS production in the presence of jute paralleled bacterial cellulase activity. High performance liquid chromatography (HPLC), matrix assisted LASER desorption/ionization-time of flight (MALDI-ToF) mass spectroscopy, and fourier transform infrared (FT-IR) spectroscopy demonstrated that the EPS synthesized in jute culture (JC) differed from that synthesized in glucose mineral salts medium (GMSM). While fucose was only a minor constituent (4.9 wt.%) of EPS from GMSM, it a major component (41.9 wt.%) of EPS synthesized in JC. This study establishes jute as an effective fermentation substrate for EPS production by a cellulase-producing bacterium.


Subject(s)
Bacillus megaterium/metabolism , Corchorus/metabolism , Extracellular Space/metabolism , Polysaccharides, Bacterial/biosynthesis , Bacillus megaterium/enzymology , Bacillus megaterium/growth & development , Cellulase/metabolism , Monosaccharides/analysis
4.
Int J Biol Macromol ; 48(4): 705-12, 2011 May 01.
Article in English | MEDLINE | ID: mdl-21382404

ABSTRACT

Bacillus pumilus UW-02, an isolate from agricultural soil irrigated with waste water was found to produce a carbohydrate polymer in the form of extracellular polysaccharide (EPS) in glucose mineral salts medium (GMSM). The recovery rates of EPS by ion-exchange and gel filtration chromatography were around 63% and 90%, respectively. As evident from HPLC and FT-IR analyses, the EPS was found to be a heteropolymer consisting glucose, mannose, xylose, arabinose, and N-acetyl glucosamine as monomer units. Different oligosaccharide combinations namely hexose(4), hexose(6) pentose(1) and hexose(10) pentose(1) are obtained after partial hydrolysis of EPS using MALDI-ToF-MS. Electron micrographs portrayed the intense affinity of the EPS molecules for each other, thereby justifying its viscosifying and thickening properties. The EPS with an average molecular weight of 218 kDa and thermal stability up to 180 °C showed pseudoplastic rheology and significant emulsifying activities.


Subject(s)
Bacillus/chemistry , Emulsifying Agents/chemistry , Polysaccharides, Bacterial/chemistry , Soil Microbiology , Chromatography, High Pressure Liquid , Emulsifying Agents/isolation & purification , Microscopy, Electron , Polysaccharides, Bacterial/isolation & purification , Rheology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Spectroscopy, Fourier Transform Infrared , Viscosity
5.
Bioresour Technol ; 101(9): 3182-7, 2010 May.
Article in English | MEDLINE | ID: mdl-20074944

ABSTRACT

The physico-chemical properties of jute fibers treated with alkali (NaOH) solution have been investigated in this study. The treatments were applied under ambient and elevated temperatures and high pressure steaming conditions. To the knowledge of these authors the influence of alkali-steam treatment on the uniaxial tensile strength of natural ligno-cellulosic fibers, such as jute, has not been investigated earlier. The results from this investigation indicate that a 30 min dipping of the fibers in 0.5% alkali solution followed by 30 min alkali-steam treatment leads to an increase in the tensile strength of up to 65%. The increase appears to be due to fiber separation and removal of non-cellulosic materials, which, in turn, resulted in an increased crystallinity.


Subject(s)
Alkalies/pharmacology , Corchorus/chemistry , Corchorus/drug effects , Lignin/chemistry , Materials Testing/methods , Steam , Tensile Strength/drug effects , Crystallization , Materials Testing/economics , Microscopy, Electron, Scanning , Spectroscopy, Fourier Transform Infrared , Surface Properties/drug effects , Temperature , X-Ray Diffraction
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