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Biomedical subjects

Munia Ganguli

Publications and source records attributed to Munia Ganguli.

8 recordsLinked to original sources

Blood compatibility of novel water soluble hyperbranched polyglycerol-based multivalent cationic polymers and their interaction with DNA.

A novel class of hyperbranched polymers based on polyglycerol (PG) and poly(ethylene glycol) (PEG) are synthesized by multibranching anionic ring opening polymerization. Multivalent cationic sites are added to these polymers by a post-amination and quarternization reactions. Blood compatibility studies using these polymers at different concentrations showed insignificant effects on complement activation, platelet activation, coagulation, erythrocyte aggregation and hemolysis compared to branched cationic polyethyleneimine (PEI). The degree of quarternization does not have large influence on the blood compatibility of the new polymers. Cytotoxicity of these polymers is significantly lower than that of PEI and is a function of quarternized nitrogen present in the polymer. Also, these polymers bind DNA in the nanomolar range and are able to condense DNA to highly compact, stable, water soluble nanoparticles in the range of 60-80 nm. Gel electrophoresis studies showed that they form electroneutral complexes with DNA around N/P ratio 1 irrespective of the percentage of quarternization under the conditions studied.

Biocompatible Materials↗

PEI-alginate nanocomposites as efficient in vitro gene transfection agents.

The positive charge on PEI was partially shielded by forming ionic nanocomposites with a polysaccharide, alginic acid, in aqueous solution, bypassing tedious chemical synthesis. The content of alginic acid was varied systematically to obtain a series of nanocomposites. The nanocomposites were first characterized by assessing the surface charge (zeta potential), size (DLS) and morphology (AFM) followed by evaluation for their DNA interaction ability, cytotoxicity and transfection efficiency on various cell lines. The transfection efficiency of PEI-alginate (6.26%) nanocomposites improved dramatically (2-16-fold over native PEI) in all the cell lines studied. However, a decrease in transfection efficiency was observed on deviating from this optimal concentration of alginic acid in nanocomposites. Cytotoxicity of PEI-alginate/DNA complexes was nearly abolished on increasing the concentration of alginic acid in nanocomposites. PEI-alginate (6.26%) nanocomposites also delivered SiRNAs efficiently into mammalian cells, resulting in 80% suppression of GFP expression. The cellular uptake and endosomal escape of PEI-alginate nanocomposites and PEI were found to follow a similar route when transfection was carried out in presence of chloroquine, bafilomycin A1, cytochalasin B and methyl-beta-cyclodextrin. The results demonstrate a versatile vector that can be used for efficient cytoplasmic delivery of a broad range of nucleic acids.

Alginates↗

Water soluble nanoparticles from PEG-based cationic hyperbranched polymer and RNA that protect RNA from enzymatic degradation.

Recent advances in understanding biological systems have proven that RNA is not merely the carrier of genetic information, but also a key molecule in regulation of gene expression and other crucial metabolic processes. Therefore, it is being considered as an ideal therapeutic candidate both for metabolic and genetic disorders. However, research involving RNA molecules faces a practical limitation since RNA is highly labile. We have developed a novel method to protect RNA from cleavage by complexing it with a hyperbranched cationic polymer. It was found that total cellular RNA isolated from yeast spontaneously interacts with the positively charged polymer to form a spherical nanoparticle morphology. This interaction protects the RNA against enzymatic degradation. This methodology can be easily adapted for long-term storage of RNA, long distance transfer of RNA, and genetic engineering using RNA as a building block.

Biopolymers↗

Nanoparticle formation from poly(acrylic acid) and oppositely charged peptides.

Cationic peptides self assemble upon interacting with sodium salt of oppositely charged polymer, poly(acrylic acid), PAA, giving rise to water-soluble nanoparticles at very low concentration (0.1 mM of PAA). The morphology of these kinds of nanoparticles is mainly governed by the composition of the complexes, which can be expressed as Z+/-, i.e., the ratio of positively charged units to the concentration of anionic units of the polymers present in the system. In the present study, at lower Z+/-, the particles are elongated in shape but adopt spherical shape of 75-100 nm in diameter at higher Z+/- values. We propose that the nanoparticles containing cationic peptides obtained by this methodology can serve as delivery system to enhance the antinociception effect of the chimeric peptide with previously administered doses.

Acrylic Resins↗

Imaging and analysis of Bacillus anthracis spore germination.

External and internal changes occurring during the process of germination of Bacillus anthracis spores were observed through atomic force microscopy (AFM) and transmission electron microscopy (TEM), respectively. AFM studies showed that in response to L-alanine (4 mM), as a germinant, the spore germinates into a vegetative cell in 3 hours. The temporal size changes occurring during the germination were gradual but the major change in size was observed between the second and third hour. TEM of spores showed the presence of varied layers, which is in accordance with previous studies. However, the integrity of these layers was lost gradually during the process of germination. The inner spore membrane remains intact even until late stages of germination, whereas the coat, outer spore membrane, and the cortical layers are discarded at the second-hour stage. The results indicate that sequential changes during the germination of a B. anthracis spore are similar to other species of the Bacillus group.

Bacillus anthracis↗

Complex formation between cationically modified gold nanoparticles and DNA: an atomic force microscopic study.

Atomic force microscopy has been used for direct visualization of the wrapping of DNA around 30-nm-sized functionalized gold nanoparticles for the first time. The morphology of the complexes seems to be dictated by the relative concentration of the nanoparticles and DNA. A higher concentration of the former leads to the formation of a network of nanoparticles assembled on DNA. This assembly pattern seems to be significantly different from the manner in which cationically modified gold nanoparticles of smaller size (< 5 nm) arrange linearly on DNA, as shown in the literature. A DNA-gold nanoparticle can be developed as a model system for in vitro studies on the mechanism of DNA condensation and also for developing novel methods of nanoparticle self-assembly on the DNA template.

Cations↗

Complexes of poly(ethylene glycol)-based cationic random copolymer and calf thymus DNA: a complete biophysical characterization.

Complete biophysical characterization of complexes (polyplexes) of cationic polymers and DNA is needed to understand the mechanism underlying nonviral therapeutic gene transfer. In this article, we propose a new series of synthesized random cationic polymers (RCPs) from methoxy poly(ethylene glycol) monomethacrylate (MePEGMA) and (3-(methacryloylamino)propyl)trimethylammonium chloride with different mole ratios (32:68, 11:89, and 6:94) which could be used as a model system to address and answer the basic questions relating to the mechanism of the interaction of calf thymus DNA (CT-DNA) and cationic polymers. The solubility of the complexes of CT-DNA and RCP was followed by turbidity measurements. It has been observed that complexes of RCP with 68 mol % MePEGMA precipitate near the charge neutralization point, whereas complexes of the other two polymers are water-soluble and stable at all compositions. Dnase 1 digestion experiments show that DNA is inaccessible when it forms complexes with RCP. Ethidium bromide exclusion and gel electrophoretic mobility show that both polymers are capable of binding with CT-DNA. Atomic force microscopy images in conjunction with light scattering experiments showed that the complexes are spherical in nature and 75-100 nm in diameter. Circular dichroism spectroscopy studies indicated that the secondary structure of DNA in the complexes is not perturbed due to the presence of poly(ethylene glycol) segments in the polymer. Furthermore, we used a combination of spectroscopic and calorimetric techniques to determine complete thermodynamic profiles accompanying the helix-coil transition of CT-DNA in the complexes. UV and differential scanning calorimetry melting experiments revealed that DNA in the complexes is more stable than in the free state and the extent of stability depends on the polymer composition. Isothermal titration calorimetry experiments showed that the binding of these RCPs to CT-DNA is associated with small exothermic enthalpy changes. A complete thermodynamic profile showed that the RCP/DNA complex formation is entropically favorable. Much broader opportunities to vary the architecture of the polymers studied here make these systems promising in addressing various basic and practical problems in gene delivery systems.

Animals↗

Nanoparticles from cationic copolymer and DNA that are soluble and stable in common organic solvents.

DNA by virtue of its superlative ability to self-assemble has found use beyond biological research in the design and fabrication of nanomaterials. However, developing novel DNA-based materials for chemical applications might be restricted due to the insoluble nature of DNA in most common organic solvents. In this Communication, we are reporting the first demonstration of making DNA soluble in a variety of nonbiological solvents such as acetonitrile, benzene, dimethyl sulfoxide (DMSO), and tetrahydrofuran with the help of poly(ethylene glycol) (PEG)-based cationic random copolymers. Because of complex formation between cationic copolymer and anionic DNA, nanoparticles are formed. These nanoparticles are expected to exhibit micelle-like structures with a nanometric core of cationic units neutralized by phosphate anions of DNA, surrounded by a shell of PEG segments. As PEG is soluble in the organic solvents used in this study, nanoparticles are stable in these solvents, making entrapped DNA soluble in these organic solvents.

Ammonium Chloride↗