Priya Vashisth

Rapid efficient synthesis and characterization of silver, gold, and bimetallic nanoparticles from the medicinal plant Plumbago zeylanica and their application in biofilm control

Background Nanoparticles (NPs) have gained significance in medical fields due to their high surface-area-to-volume ratio. In this study, we synthesized NPs from a medicinally important plant – Plumbago zeylanica. Materials and methods Aqueous root extract of P. zeylanica (PZRE) was analyzed for the presence of flavonoids, sugars, and organic acids using high-performance thin-layer chromatography (HPTLC), gas chromatography-time of flight-mass spectrometry (GC-TOF-MS), and biochemical methods. The silver NPs (AgNPs), gold NPs (AuNPs), and bimetallic NPs (AgAuNPs) were synthesized from root extract and characterized using ultraviolet-visible spectra, X-ray diffraction (XRD), energy-dispersive spectrometry (EDS), transmission electron microscopy (TEM), and dynamic light scattering (DLS). The effects of these NPs on Acinetobacter baumannii, Staphylococcus aureus, and Escherichia coli biofilms were studied using quantitative biofilm inhibition and disruption assays, as well as using fluorescence, scanning electron microscopy, and atomic force microscopy. Results PZRE showed the presence of phenolics, such as plumbagin, and flavonoids, in addition to citric acid, sucrose, glucose, fructose, and starch, using HPTLC, GC-TOF-MS, and quantitative analysis. Bioreduction of silver nitrate (AgNO3) and chloroauric acid (HAuCl4) were confirmed at absorbances of 440 nm (AgNPs), 570 nm (AuNPs), and 540 nm (AgAuNPs), respectively. The maximum rate of synthesis at 50°C was achieved with 5 mM AgNO3 within 4.5 hours for AgNPs; and with 0.7 mM HAuCl4 within 5 hours for AuNPs. The synthesis of AgAuNPs, which completed within 90 minutes with 0.7 mM AgNO3 and HAuCl4, was found to be the fastest. Fourier-transform infrared spectroscopy confirmed bioreduction, while EDS and XRD patterns confirmed purity and the crystalline nature of the NPs, respectively. TEM micrographs and DLS showed about 60 nm monodispersed Ag nanospheres, 20–30 nm Au nanospheres adhering to form Au nanotriangles, and about 90 nm hexagonal blunt-ended AgAuNPs. These NPs also showed antimicrobial and antibiofilm activity against E. coli, A. baumannii, S. aureus, and a mixed culture of A. baumannii and S. aureus. AgNPs inhibited biofilm in the range of 96%–99% and AgAuNPs from 93% to 98% in single-culture biofilms. AuNPs also showed biofilm inhibition, with the highest of 98% in S. aureus. AgNPs also showed good biofilm disruption, with the highest of 88% in A. baumannii. Conclusion This is the first report on rapid and efficient synthesis of AgNPs, AuNPs and AgAuNPs from P. zeylanica and their effect on quantitative inhibition and disruption of bacterial biofilms.

Antibiofilm activity of quercetin- encapsulated cytocompatible nanofibers against Candida albicans

In this study, nanofibers against pro dimorphic fungal sessile growth were developed. Quercetin was successfully encapsulated within poly(d,l-lactide-co-glycolide)–poly(ε-caprolactone) nanofibers using an electrospinning technique. Field emission scanning electron microscopy, fluorescent microscopy, and Fourier-transformed infrared spectrometer were used to confirm the formation as well as encapsulation of quercetin within the nanofibers. These fabricated nanofibers were further evaluated to determine the effectiveness of the antibiofilm activity against Candida albicans. The cytocompatibility of quercetin-encapsulated nanofibers was found to be similar to control and pure polymeric nanofibers based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay against human embryonic kidney (HEK-293) cell lines. These fabricated nanofibers potentially could be used as coatings on biomedical devices to inhibit microbial contaminations.

Process optimization for fabrication of gellan based electrospun nanofibers.

In this investigation, the nanofiber formation ability of gellan, a FDA approved low cost natural polysaccharide, has been achieved for the first time using electrospinning technique. The gellan based ultrafine nanofibers were fabricated by using a blend mixture of gellan with another biodegradable polymer polyvinyl alcohol (PVA). The morphology of resulting gellan-PVA nanofibers was analyzed using field emission scanning electron microscopy (FESEM). The mass ratio of 50:50 for gellan:PVA was recorded as an optimum solution ratio to obtain uniform bead free nanofibers with an average diameter of 40 ± 15.8 nm. Data depicted that among different parameters evaluated, viscosity and the mass ratio of gellan:PVA were the key parameters that influence the nanofiber morphology and diameter.

Synthesis and characterization of crosslinked gellan/PVA nanofibers for tissue engineering application

Electrospun nanofibers based on gellan are considered as promising biomaterial for tissue engineering and wound healing applications. However, major hurdles in usage of these nanofibers are their poor stability and deprived structural consistency in aqueous medium which is a prerequisite for their application in the biomedical sector. In this investigation, three dimensional nanofibers, consisting of gellan and PVA have been fabricated and then stabilized under various crosslinking conditions in order to improve their physiochemical stability. The impacts of different crosslinking procedures on the gellan/PVA nanofibers were examined in terms of changes in morphological, mechanical, swelling and biological properties. Superior tensile strength and strain was recorded in case of crosslinked nanofibers as compared to non-crosslinked nanofibers. Contact angles and swelling properties of fabricated gellan/PVA nanofibers were found to vary with the crosslinking method. All crosslinking conditions were evaluated with regard to their response towards human dermal fibroblast (3T3L1) cells. Biocompatibility studies suggested that the fabricated crosslinked gellan/PVA nanofibers hold a great prospective in the biomedical engineering arena.

A novel gellan-PVA nanofibrous scaffold for skin tissue regeneration: Fabrication and characterization.

In this investigation, we have introduced novel electrospun gellan based nanofibers as a hydrophilic scaffolding material for skin tissue regeneration. These nanofibers were fabricated using a blend mixture of gellan with polyvinyl alcohol (PVA). PVA reduced the repulsive force of resulting solution and lead to formation of uniform fibers with improved nanostructure. Field emission scanning electron microscopy (FESEM) confirmed the average diameter of nanofibers down to 50 nm. The infrared spectra (IR), differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis evaluated the crosslinking, thermal stability and highly crystalline nature of gellan-PVA nanofibers, respectively. Furthermore, the cell culture studies using human dermal fibroblast (3T3L1) cells established that these gellan based nanofibrous scaffold could induce improved cell adhesion and enhanced cell growth than conventionally proposed gellan based hydrogels and dry films. Importantly, the nanofibrous scaffold are biodegradable and could be potentially used as a temporary substrate/or biomedical graft to induce skin tissue regeneration.

Biomedical applications of ferulic acid encapsulated electrospun nanofibers

Ferulic acid is a ubiquitous phytochemical that holds enormous therapeutic potential but has not gained much consideration in biomedical sector due to its less bioavailability, poor aqueous solubility and physiochemical instability. In present investigation, the shortcomings associated with agro-waste derived ferulic acid were addressed by encapsulating it in electrospun nanofibrous matrix of poly (d,l-lactide-co-glycolide)/polyethylene oxide. Fluorescent microscopic analysis revealed that ferulic acid predominantly resides in the core of PLGA/PEO nanofibers. The average diameters of the PLGA/PEO and ferulic acid encapsulated PLGA/PEO nanofibers were recorded as 125 ± 65.5 nm and 150 ± 79.0 nm, respectively. The physiochemical properties of fabricated nanofibers are elucidated by IR, DSC and NMR studies. Free radical scavenging activity of fabricated nanofibers were estimated using di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH) assay. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay confirmed the cytotoxicity of ferulic acid encapsulated nanofibers against hepatocellular carcinoma (HepG2) cells. These ferulic acid encapsulated nanofibers could be potentially explored for therapeutic usage in biomedical sector.

Drug functionalized microbial polysaccharide based nanofibers as transdermal substitute

In order to promote the natural healing process, drug-functionalized nanofibrous transdermal substitute was fabricated using gellan as chief polymer and polyvinyl alcohol (PVA) as supporting polymer via electrospinning technique. These fabricated nanofibers physiochemically mimic the extracellular matrix (ECM) which supports the cell growth. For neo-tissue regeneration in a sterilized environment, amoxicillin (Amx) was entrapped within these nanofibers. Entrapment of Amx in the nanofibers was confirmed by FESEM, FTIR, XRD and TG analysis. In vitro cell culture studies revealed that the fabricated non-cytotoxic nanofibers promoted enhance cell adherence and proliferation of human keratinocytes. A preliminary in vivo study performed on rat model for full thickness skin excision wound demonstrated the prompt re-epithelialization in early phase and quicker collagen deposition in later phases of wound healing in case of Amx-functionalized gellan/PVA nanofibers. Data collectively confirmed the potential usage of gellan based electrospun nanofibers as transdermal substitute for faster skin restoration.

Ofloxacin loaded gellan/PVA nanofibers - Synthesis, characterization and evaluation of their gastroretentive/mucoadhesive drug delivery potential

The purpose of this investigation is to formulate a gastroretentive sustained drug release system for ofloxacin to improve its retention time, pharmacological activity, bioavailability and therapeutic efficacy in the stomach. Ofloxacin loaded gellan/poly vinyl alcohol (PVA) nanofibers were fabricated using a simple and versatile electrospinning technique. The fabricated nanofibers were evaluated for percent drug encapsulation efficiency and in vitro drug release in simulated gastric medium (pH1.2). The in vitro release profile and kinetic studies for drug indicated the sustained release of ofloxacin from the nanofibers through Fickian diffusion kinetics. The antimicrobial activity of the ofloxacin loaded nanofibers was assessed in comparison to the pure ofloxacin by means of minimal inhibitory concentrations (MIC) against microbial strains of Enterococcus faecalis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. The optimized ofloxacin loaded gellan/PVA nanofibers displayed biphasic drug release profile with considerable mucoadhesion and gastric retention in the rats gastric mucosal membrane. Data obtained, suggested that the developed gastroretentive drug delivery can potentially enhance the pharmacological activity of ofloxacin and can also serve as a viable alternative for improving drug bioavailability via oral route.

A controlled release system for quercetin from biodegradable poly(lactide-co-glycolide)–polycaprolactone nanofibers and its in vitro antitumor activity

Quercetin is a potent natural antioxidant but has limited therapeutic applications due to its short half-life in body fluids. In order to improve the efficacy of quercetin and overcome its shortcomings, quercetin-encapsulated electrospun poly(lactic-co-glycolic acid)–poly(ε-caprolactone) nanofibrous controlled release system was developed using electrospinning technique. Fourier transform infrared spectroscopy, thermogravimetric, and X-ray diffraction analysis suggested the incorporation, thermal stability, and existence of encapsulated quercetin in semicrystalline state in the nanofibers. The release profiles of quercetin from the poly(lactide-co-glycolide)–polycaprolactone nanofibers in phosphate-buffered saline showed controlled release of quercetin up to 120 h. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed an evident inhibition effect of quercetin-encapsulated nanofibers against human hepatocellular carcinoma (HepG2), and the inhibition rate of 29%, 72%, and 80.1% were recorded for 1%, 2%, and 4% quercetin-encapsulated nanofibers, respectively. The formulated drug delivery system could be potentially used as an implantable anticancer drug in clinical applications in the future.

Synergistic effects of Woodfordia fruticosa gold nanoparticles in preventing microbial adhesion and accelerating wound healing in Wistar albino rats in vivo

Therapeutic effectiveness of biogenically synthesized Woodfordia fruticosa nano-gold particles (WfAuNPs) has been claimed in this study which prevents microbial adhesion and enhanced wound healing potential on Wistar albino rats. The synthesized WfAuNPs were characterized using several biophysical techniques such as UV-Visible Spectroscopy (UV-vis), X-Ray Diffraction (XRD), Dynamic Light Scattering (DLS), Zeta Potential, Fourier Transform Infrared Spectroscopy (FTIR), Field Emission Scanning Electron Microscopy (FE-SEM), Atomic Force Microscopy (AFM) and High Resolution Transmission Electron Microscopy (HR-TEM) analysis. The synthesized WfAuNPs in the size range of 10-20nm were used to develop 1% Carbopol® 934 based nano gold formulation (WfAuNPs-Carbopol® 934). The WfAuNPs-Carbopol® 934 nanoformulation was evaluated using viscosity and spreadability measurements. The wound healing potential of WfAuNPs-Carbopol® 934 monitored up to 12days was confirmed by performing wound contraction (%), epithelialization, and histopathological studies done in vivo on Wistar albino rats. The hydroxyproline content was also measured in the re-epithelized skin for quantification of collagen content. The effects of WfAuNPs on microbial adhesion leading to biofilm formation were evaluated against Candida albicans and Cryptococcus neoformans fungal strains. The respective Minimum Inhibitory Concentration (MIC80), Biofilm Inhibitory Concentration (BIC80) and Biofilm Eradication Concentration (BEC80) values of C. albicans was found to be 16, 32, 256μg/ml respectively while for C. neoformans it was recorded to be 32, 64, 256μg/ml respectively. Data obtained, confirmed the effectiveness in preventing microbial adhesion and wound healing potential of the WfAuNPs as compared to current marketed formulations.