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Biomedical subjects

Shanta Raj Bhattarai

Publications and source records attributed to Shanta Raj Bhattarai.

7 recordsLinked to original sources

Novel block copolymer (PPDO/PLLA-b-PEG): enhancement of DNA uptake and cell transfection.

The cationic lipid mediated uptake of plasmid DNA by cells in monolayer culture was significantly enhanced with an aqueous solution of the block copolymer poly(p-dioxanone-co-L-lactide)-b-poly(ethylene glycol) (PPDO/PLLA-b-PEG). Plasmid uptake studies with DNA encoding the beta-galactosidase gene and cytotoxicity evaluations were performed on MCF-7, NIH 3T3 and CT-26 cell lines. Transfection yields and time courses for maximum release of FITC labeled DNA in MCF-7 cells were observed and quantified by beta-galactosidase assay and spectrofluorometry, respectively. The reported results suggest that the studied block copolymer might be useful for the enhancement of polycation mediated transfection and could find application in gene therapy.

3T3 Cells↗

Hydrophilic nanofibrous structure of polylactide; fabrication and cell affinity.

Microstructure and architecture of the scaffolds along with the surface chemistry exert profound effect on biological activity (cell distribution, proliferation, and differentiation). For the biological activity, scaffolds in tissue engineering have been widely designed. The objective of this study was to develop hydrophilic nanofibrous structure of polylactides (PLLA) polymer in the form of nonwoven mat by electrospinning technique, and further evaluate the fibroblast NIH3T3 cell proliferation, morphology, and cell-matrix interaction. Hydrophilicity of the PLLA fibers was improved by adding small fraction of low molecular weight polyethylene glycol (PEG) into the electrospinning solution. Four different ratio types (100/0, 80/20, 70/30, and 50/50) of PLLA/PEG electrospun matrices were fabricated, and the pore characteristics, tensile properties, contact angle, and hydrolytic degradation were observed. Furthermore, scanning electron microscope (SEM) and fluorescence actin staining images were used for micro-observation of cell-matrix interaction and cell morphology. It was found that the electrospun mat of PLLA/PEG (80/20), composed of fibers with diameters in the range 540-850 nm, majority of pore diameter less than 100 microm, tensile strength 8 MPa, elongation 150%, porosity more than 90%, and improved hydrophilicity with slow hydrolytic degradation, is favorable for biological activity of NIH3T3 fibroblast cell. Based on these results, the correct composition of PLLA and PEG in the porous electrospun matrix (i.e., PLLA/PEG (80/20)) will be a better candidate rather than other compositions of PLLA/PEG as well as hydrophobic PLLA for application in tissue engineering.

3T3 Cells↗

Stabilization of gold nanoparticles by hydrophobically-modified polycations.

Surface-modified gold nanoparticles have pronounced benefits in the biomedical field due to their significant interaction with delivery materials. In the present study we used hydrophobically-modified polycations (i.e., N-acylated chitosan) to stabilize gold nanoparticles. Aliphatic hydrophobic groups, having carbon chains of different lengths, were first grafted onto the backbone of chitosan by N-acylation with fatty-acid chlorides in order to increase its hydrophobicity. Gold nanoparticles stabilized with native chitosan and N-acylated chitosan were prepared by the graft-onto approach. Chemical modification and its quantification were studied by Fourier-transform infrared (FT-IR) spectroscopy. Further, the stabilized gold nanoparticles were characterized by different physico-chemical techniques such as UV-Vis, FT-IR, TEM, TGA and DLS. Spectral studies of gold nanoparticles show the backbone and the side chain functional groups of chitosan were not cleaved during the conjugation process. TEM observations revealed that the modified chitosan gold nanoparticles were well dispersed and spherical in shape with average size around 10-12 nm in triply-distilled water at pH 7.4, whereas the native chitosan gold nanoparticles appeared as clusters with 9.9 nm as average diameter and were dispersed only in dilute HCl. The size of modified chitosan gold nanoparticles varied depending on the length of grafting molecules.

Biocompatible Materials↗

Deposition of gold nanoparticles on electrospun MgTiO3 ceramic nanofibers.

A simple method to deposit spherical gold nanoparticles on the surface of MgTiO3 ceramic nanofibers is presented. Electrospun MgTiO3/poly(vinyl acetate) (PVAc) hybrid nanofibers were calcined at 650 degrees C to obtain phase pure ceramic MgTiO3 nanofibers with 100-150 nm diameters. These ceramic nanofibers were immersed in an aqueous solution of HAuCl4 containing poly(vinyl alcohol) (PVA) as capping agent followed by photoreduction at 365 nm to get a novel Au-MgTiO3 nanocomposite. The formation of gold nanoparticles upon irradiation was confirmed by the appearance of a surface plasmon band (SPB) at 590 nm in the UV-visible absorption spectra. The surface morphology and elemental compositions were analyzed by the scanning electron microscope (SEM) equipped with energy dispersive X-ray (EDX), and transmission electron microscope (TEM). X-ray diffraction (XRD) and selected area diffraction (SAED) pattern in TEM revealed the crystallization of gold by exhibiting strong diffractions correspond to Au(111) and Au(200) crystalline planes in addition to the MgTiO3 diffraction.

Ceramics↗

Novel fabricated matrix via electrospinning for tissue engineering.

Electric field-driven fiber formation (electrospinning) is developing into a practical means for preparing novel porous filament with unusual structures and affordable mechanical properties. Polycaprolactone (PCL) was dissolved in solvent mixtures of methylene chloride/N,N-dimethyl formamide with ratios of 100/0, 75/25, and 50/50 (v/v) for electrospinning. The filament was formed by coagulation of the spinning solution following the well-known principle of phase separation in polymer solutions valid in other wet shaping processes. A strand of electrospun porous filament consisted of fibers ranging from 0.5 to 12 microm in diameter. To evaluate the feasibility of three-dimensional fabric as scaffold matrices, the plain weave, which is the simplest of the weaves and the most common, was prepared with porous PCL filament. The growth characteristics of MCF-7 mammary carcinoma cells in the woven fabrics showed the important role of matrix microstructure in proliferation. This study has shown that woven fabrics, consisting of porous filaments via electrospinning, may be suitable candidates as tissue engineering scaffolds.

Carcinoma↗

Novel biodegradable electrospun membrane: scaffold for tissue engineering.

Nonwoven fibrous matrixes have been widely used as scaffolds in tissue engineering, and modification of microstructure of these matrices is needed to organize cells in three-dimensional space with spatially balanced proliferation and differentiation required for functional tissue development. The objective of this study was fabrication of nanofibrous matrix from novel biodegradable poly(p-dioxanone-co-L-lactide)-block-poly(ethylene glycol) (PPDO/PLLA-b-PEG) copolymer, and to examine cell proliferation, morphology of cell-matrix interaction with the electrospun nanofibrous matrix. The electrospun structure composed of PPDO/PLLA-b-PEG fibers with an average diameters of 380 nm, median pore size 8 microm, porosity more than 80% and mechanical strength 1.4 MPa, is favorable for cell-matrix interaction and supports the active biocompatibility of the structure. NIH 3T3 fibroblast cell seeded on the structure tend to maintain phenotypic shape and guided growth according to nanofiber orientation. Good capability of the nanofibrous structure for supporting the cell attachment and proliferation are observed. This novel biodegradable scaffold will be applicable for tissue engineering based upon its unique architecture, which acts to support and guide cell growth.

3T3 Cells↗

Novel polymeric micelles of amphiphilic triblock copolymer poly (p-dioxanone-co-L-lactide)-block-poly (ethylene glycol).

PURPOSE: The objective of this study is to characterize the micelles of novel block copolymer of poly (p-Dioxanone-co-L-Lactide)-block-Poly (ethylene glycol) (PPDO/PLLA-b-PEG-) and evaluate its ability to induce gene transfection. METHODS: The ability of the block copolymer to self-assemble was determined by viscometery, dye solublization, NMR spectra and dynamic light scattering. The Trypan blue assay for in vitro biocompatibility of the block copolymer was carried out with NIH 3T3, CT-26 and MCF-7 cells, and beta-glactosidase assay was applied to measure the transfection efficiency of the block copolymer on MCF-7 breast cancer cell. RESULTS: Depending on the block lengths and molecular weights, solubility of the polymeric samples in water was varied. Diluted aqueous solution properties of the copolymer were studied. 1,6-Diphenyl-1,3,5-hexatriene solubilization and 1H NMR spectra carried out in CDCl3 and D2O, were used to prove the existence of hydrophobic domains as the core of micelle. Average particle size of 60-165 nm with low polydispersity, and lower negative zeta potential of -3 to -14 mV were observed on the aqueous copolymer dispersion. Copolymer was found with almost no cytotoxic effect and was able to promote the transfection efficiency (about 3-fold) in MCF-7 cells. CONCLUSIONS: The PPDO/PLLA-b-PEG copolymer has ability to assemble into nanoscopic structures in aqueous environment, which enable to enhance gene transfection.

Animals↗